EV-TRACK

Search > Results

You searched for: EV230812 (EV-TRACK ID)

Showing 1 - 14 of 14

Results list Study Tree

Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV230812	2/14	Streptomyces sp. Mg1	PSK0558	(d)(U)C DG Filtration	Hoefler BC	2017	71%
Study summary Full title A Link between Linearmycin Biosynthesis and Extracellular Vesicle Genesis Connects Specialized Metabolism and Bacterial Membrane Physiology. All authors Hoefler BC, Stubbendieck RM, Josyula NK, Moisan SM, Schulze EM, Straight PD Journal Cell Chem Biol Abstract Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and (show more...)Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and metal scavengers. Little is known about how specialized metabolites are processed and trafficked for their diverse competitive functions. Linearmycins A and B are linear polyketides with antifungal and antibacterial activity but are colony-localized in imaging mass spectrometry of Streptomyces sp. Mg1 (S. sp. Mg1). To decipher a connection between colony localization and antibiotic activity, we identified the linearmycin gene cluster and investigated linearmycin production and distribution by S. sp. Mg1. Our results uncover a large family of variant linearmycins with limited solubility in aqueous solution. We hypothesized that extracellular vesicles may traffic the lipid-like linearmycins. We found that vesicles isolated from culture supernatants contained linearmycins. Surprisingly, abolishing production of linearmycins in S. sp. Mg1 also diminished extracellular vesicle production. Our results reveal integration of linearmycin biosynthesis with production of extracellular vesicles, suggesting a deep connection between specialized metabolism and bacterial membrane physiology. (hide) EV-METRIC 71% (96th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin ΔlnyI Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Density gradient Filtration Protein markers EV: None non-EV: None Proteomics yes EV density (g/ml) not specified Show all info Study aim Biogenesis/cargo sorting/Identification of content (omics approaches) Sample Species Streptomyces sp. Mg1 Sample Type Cell culture supernatant EV-producing cells PSK0558 EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 10,000 g and 50,000 g Equal to or above 150,000 g Pelleting performed Yes Pelleting: rotor type Type 45 Ti Pelleting: speed (g) 235000 Density gradient Type Discontinuous Number of initial discontinuous layers 8 Lowest density fraction 10% Highest density fraction 50% Total gradient volume, incl. sample (mL) 14 Sample volume (mL) 1 Orientation Bottom-up Speed (g) 100400 Duration (min) 960 Fraction volume (mL) 1 Fraction processing Centrifugation Pelleting: volume per fraction 4mL Pelleting: speed (g) 120000 Pelleting: adjusted k-factor 1.857 Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method Not determined Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis DLS EM EM-type Transmission-EM Image type Wide-field EV concentration Yes

	EV230812	3/14	Streptomyces sp. Mg1	PSK0558	(d)(U)C DG Filtration	Hoefler BC	2017	71%
Study summary Full title A Link between Linearmycin Biosynthesis and Extracellular Vesicle Genesis Connects Specialized Metabolism and Bacterial Membrane Physiology. All authors Hoefler BC, Stubbendieck RM, Josyula NK, Moisan SM, Schulze EM, Straight PD Journal Cell Chem Biol Abstract Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and (show more...)Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and metal scavengers. Little is known about how specialized metabolites are processed and trafficked for their diverse competitive functions. Linearmycins A and B are linear polyketides with antifungal and antibacterial activity but are colony-localized in imaging mass spectrometry of Streptomyces sp. Mg1 (S. sp. Mg1). To decipher a connection between colony localization and antibiotic activity, we identified the linearmycin gene cluster and investigated linearmycin production and distribution by S. sp. Mg1. Our results uncover a large family of variant linearmycins with limited solubility in aqueous solution. We hypothesized that extracellular vesicles may traffic the lipid-like linearmycins. We found that vesicles isolated from culture supernatants contained linearmycins. Surprisingly, abolishing production of linearmycins in S. sp. Mg1 also diminished extracellular vesicle production. Our results reveal integration of linearmycin biosynthesis with production of extracellular vesicles, suggesting a deep connection between specialized metabolism and bacterial membrane physiology. (hide) EV-METRIC 71% (96th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin ΔlnyI + lnyI (complement) Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Density gradient Filtration Protein markers EV: None non-EV: None Proteomics yes EV density (g/ml) not specified Show all info Study aim Biogenesis/cargo sorting/Identification of content (omics approaches) Sample Species Streptomyces sp. Mg1 Sample Type Cell culture supernatant EV-producing cells PSK0558 EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 10,000 g and 50,000 g Equal to or above 150,000 g Pelleting performed Yes Pelleting: rotor type Type 45 Ti Pelleting: speed (g) 235000 Density gradient Type Discontinuous Number of initial discontinuous layers 8 Lowest density fraction 10% Highest density fraction 50% Total gradient volume, incl. sample (mL) 14 Sample volume (mL) 1 Orientation Bottom-up Speed (g) 100400 Duration (min) 960 Fraction volume (mL) 1 Fraction processing Centrifugation Pelleting: volume per fraction 4mL Pelleting: speed (g) 120000 Pelleting: adjusted k-factor 1.857 Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method Not determined Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis DLS EM EM-type Transmission-EM Image type Wide-field EV concentration Yes

	EV230812	1/14	Streptomyces sp. Mg1	PSK0558	(d)(U)C DG Filtration	Hoefler BC	2017	57%
Study summary Full title A Link between Linearmycin Biosynthesis and Extracellular Vesicle Genesis Connects Specialized Metabolism and Bacterial Membrane Physiology. All authors Hoefler BC, Stubbendieck RM, Josyula NK, Moisan SM, Schulze EM, Straight PD Journal Cell Chem Biol Abstract Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and (show more...)Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and metal scavengers. Little is known about how specialized metabolites are processed and trafficked for their diverse competitive functions. Linearmycins A and B are linear polyketides with antifungal and antibacterial activity but are colony-localized in imaging mass spectrometry of Streptomyces sp. Mg1 (S. sp. Mg1). To decipher a connection between colony localization and antibiotic activity, we identified the linearmycin gene cluster and investigated linearmycin production and distribution by S. sp. Mg1. Our results uncover a large family of variant linearmycins with limited solubility in aqueous solution. We hypothesized that extracellular vesicles may traffic the lipid-like linearmycins. We found that vesicles isolated from culture supernatants contained linearmycins. Surprisingly, abolishing production of linearmycins in S. sp. Mg1 also diminished extracellular vesicle production. Our results reveal integration of linearmycin biosynthesis with production of extracellular vesicles, suggesting a deep connection between specialized metabolism and bacterial membrane physiology. (hide) EV-METRIC 57% (92nd percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin Control condition Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Density gradient Filtration Protein markers EV: None non-EV: None Proteomics yes EV density (g/ml) not specified Show all info Study aim Biogenesis/cargo sorting/Identification of content (omics approaches) Sample Species Streptomyces sp. Mg1 Sample Type Cell culture supernatant EV-producing cells PSK0558 EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 10,000 g and 50,000 g Equal to or above 150,000 g Pelleting performed Yes Pelleting: rotor type Type 45 Ti Pelleting: speed (g) 235000 Density gradient Type Discontinuous Number of initial discontinuous layers 8 Lowest density fraction 10% Highest density fraction 50% Total gradient volume, incl. sample (mL) 14 Sample volume (mL) 1 Orientation Bottom-up Speed (g) 100400 Duration (min) 960 Fraction volume (mL) 1 Fraction processing Centrifugation Pelleting: volume per fraction 4mL Pelleting: speed (g) 120000 Pelleting: adjusted k-factor 1.857 Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method Not determined Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis DLS EM EM-type Transmission-EM Image type Wide-field Extra information TEM was performed for the time course experiment as well, but no images in paper

	EV230812	4/14	Streptomyces sp. Mg1	PSK0558	(d)(U)C DG Filtration	Hoefler BC	2017	57%
Study summary Full title A Link between Linearmycin Biosynthesis and Extracellular Vesicle Genesis Connects Specialized Metabolism and Bacterial Membrane Physiology. All authors Hoefler BC, Stubbendieck RM, Josyula NK, Moisan SM, Schulze EM, Straight PD Journal Cell Chem Biol Abstract Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and (show more...)Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and metal scavengers. Little is known about how specialized metabolites are processed and trafficked for their diverse competitive functions. Linearmycins A and B are linear polyketides with antifungal and antibacterial activity but are colony-localized in imaging mass spectrometry of Streptomyces sp. Mg1 (S. sp. Mg1). To decipher a connection between colony localization and antibiotic activity, we identified the linearmycin gene cluster and investigated linearmycin production and distribution by S. sp. Mg1. Our results uncover a large family of variant linearmycins with limited solubility in aqueous solution. We hypothesized that extracellular vesicles may traffic the lipid-like linearmycins. We found that vesicles isolated from culture supernatants contained linearmycins. Surprisingly, abolishing production of linearmycins in S. sp. Mg1 also diminished extracellular vesicle production. Our results reveal integration of linearmycin biosynthesis with production of extracellular vesicles, suggesting a deep connection between specialized metabolism and bacterial membrane physiology. (hide) EV-METRIC 57% (92nd percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin WT 24h Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Density gradient Filtration Protein markers EV: None non-EV: None Proteomics yes EV density (g/ml) not specified Show all info Study aim Biogenesis/cargo sorting/Identification of content (omics approaches) Sample Species Streptomyces sp. Mg1 Sample Type Cell culture supernatant EV-producing cells PSK0558 EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 10,000 g and 50,000 g Equal to or above 150,000 g Pelleting performed Yes Pelleting: rotor type Type 45 Ti Pelleting: speed (g) 235000 Density gradient Type Discontinuous Number of initial discontinuous layers 6 Lowest density fraction 0% Highest density fraction 50% Total gradient volume, incl. sample (mL) 2.1 Sample volume (mL) 0.2 Orientation Bottom-up Speed (g) 120000 Duration (min) 1020 Fraction volume (mL) 0.2 Fraction processing Centrifugation Pelleting: volume per fraction 2.1 Pelleting: speed (g) 120000 Pelleting: adjusted k-factor TDB Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method Not determined Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis None

	EV230812	5/14	Streptomyces sp. Mg1	PSK0558	(d)(U)C DG Filtration	Hoefler BC	2017	57%
Study summary Full title A Link between Linearmycin Biosynthesis and Extracellular Vesicle Genesis Connects Specialized Metabolism and Bacterial Membrane Physiology. All authors Hoefler BC, Stubbendieck RM, Josyula NK, Moisan SM, Schulze EM, Straight PD Journal Cell Chem Biol Abstract Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and (show more...)Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and metal scavengers. Little is known about how specialized metabolites are processed and trafficked for their diverse competitive functions. Linearmycins A and B are linear polyketides with antifungal and antibacterial activity but are colony-localized in imaging mass spectrometry of Streptomyces sp. Mg1 (S. sp. Mg1). To decipher a connection between colony localization and antibiotic activity, we identified the linearmycin gene cluster and investigated linearmycin production and distribution by S. sp. Mg1. Our results uncover a large family of variant linearmycins with limited solubility in aqueous solution. We hypothesized that extracellular vesicles may traffic the lipid-like linearmycins. We found that vesicles isolated from culture supernatants contained linearmycins. Surprisingly, abolishing production of linearmycins in S. sp. Mg1 also diminished extracellular vesicle production. Our results reveal integration of linearmycin biosynthesis with production of extracellular vesicles, suggesting a deep connection between specialized metabolism and bacterial membrane physiology. (hide) EV-METRIC 57% (92nd percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin WT 36h Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Density gradient Filtration Protein markers EV: None non-EV: None Proteomics yes EV density (g/ml) not specified Show all info Study aim Biogenesis/cargo sorting/Identification of content (omics approaches) Sample Species Streptomyces sp. Mg1 Sample Type Cell culture supernatant EV-producing cells PSK0558 EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 10,000 g and 50,000 g Equal to or above 150,000 g Pelleting performed Yes Pelleting: rotor type Type 45 Ti Pelleting: speed (g) 235000 Density gradient Type Discontinuous Number of initial discontinuous layers 6 Lowest density fraction 0% Highest density fraction 50% Total gradient volume, incl. sample (mL) 2.1 Sample volume (mL) 0.2 Orientation Bottom-up Speed (g) 120000 Duration (min) 1020 Fraction volume (mL) 0.2 Fraction processing Centrifugation Pelleting: volume per fraction 2.1 Pelleting: speed (g) 120000 Pelleting: adjusted k-factor TDB Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method Not determined Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis None

	EV230812	6/14	Streptomyces sp. Mg1	PSK0558	(d)(U)C DG Filtration	Hoefler BC	2017	57%
Study summary Full title A Link between Linearmycin Biosynthesis and Extracellular Vesicle Genesis Connects Specialized Metabolism and Bacterial Membrane Physiology. All authors Hoefler BC, Stubbendieck RM, Josyula NK, Moisan SM, Schulze EM, Straight PD Journal Cell Chem Biol Abstract Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and (show more...)Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and metal scavengers. Little is known about how specialized metabolites are processed and trafficked for their diverse competitive functions. Linearmycins A and B are linear polyketides with antifungal and antibacterial activity but are colony-localized in imaging mass spectrometry of Streptomyces sp. Mg1 (S. sp. Mg1). To decipher a connection between colony localization and antibiotic activity, we identified the linearmycin gene cluster and investigated linearmycin production and distribution by S. sp. Mg1. Our results uncover a large family of variant linearmycins with limited solubility in aqueous solution. We hypothesized that extracellular vesicles may traffic the lipid-like linearmycins. We found that vesicles isolated from culture supernatants contained linearmycins. Surprisingly, abolishing production of linearmycins in S. sp. Mg1 also diminished extracellular vesicle production. Our results reveal integration of linearmycin biosynthesis with production of extracellular vesicles, suggesting a deep connection between specialized metabolism and bacterial membrane physiology. (hide) EV-METRIC 57% (92nd percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin WT 48h Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Density gradient Filtration Protein markers EV: None non-EV: None Proteomics yes EV density (g/ml) not specified Show all info Study aim Biogenesis/cargo sorting/Identification of content (omics approaches) Sample Species Streptomyces sp. Mg1 Sample Type Cell culture supernatant EV-producing cells PSK0558 EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 10,000 g and 50,000 g Equal to or above 150,000 g Pelleting performed Yes Pelleting: rotor type Type 45 Ti Pelleting: speed (g) 235000 Density gradient Type Discontinuous Number of initial discontinuous layers 6 Lowest density fraction 0% Highest density fraction 50% Total gradient volume, incl. sample (mL) 2.1 Sample volume (mL) 0.2 Orientation Bottom-up Speed (g) 120000 Duration (min) 1020 Fraction volume (mL) 0.2 Fraction processing Centrifugation Pelleting: volume per fraction 2.1 Pelleting: speed (g) 120000 Pelleting: adjusted k-factor TDB Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method Not determined Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis None

	EV230812	7/14	Streptomyces sp. Mg1	PSK0558	(d)(U)C DG Filtration	Hoefler BC	2017	57%
Study summary Full title A Link between Linearmycin Biosynthesis and Extracellular Vesicle Genesis Connects Specialized Metabolism and Bacterial Membrane Physiology. All authors Hoefler BC, Stubbendieck RM, Josyula NK, Moisan SM, Schulze EM, Straight PD Journal Cell Chem Biol Abstract Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and (show more...)Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and metal scavengers. Little is known about how specialized metabolites are processed and trafficked for their diverse competitive functions. Linearmycins A and B are linear polyketides with antifungal and antibacterial activity but are colony-localized in imaging mass spectrometry of Streptomyces sp. Mg1 (S. sp. Mg1). To decipher a connection between colony localization and antibiotic activity, we identified the linearmycin gene cluster and investigated linearmycin production and distribution by S. sp. Mg1. Our results uncover a large family of variant linearmycins with limited solubility in aqueous solution. We hypothesized that extracellular vesicles may traffic the lipid-like linearmycins. We found that vesicles isolated from culture supernatants contained linearmycins. Surprisingly, abolishing production of linearmycins in S. sp. Mg1 also diminished extracellular vesicle production. Our results reveal integration of linearmycin biosynthesis with production of extracellular vesicles, suggesting a deep connection between specialized metabolism and bacterial membrane physiology. (hide) EV-METRIC 57% (92nd percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin WT 72h Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Density gradient Filtration Protein markers EV: None non-EV: None Proteomics yes EV density (g/ml) not specified Show all info Study aim Biogenesis/cargo sorting/Identification of content (omics approaches) Sample Species Streptomyces sp. Mg1 Sample Type Cell culture supernatant EV-producing cells PSK0558 EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 10,000 g and 50,000 g Equal to or above 150,000 g Pelleting performed Yes Pelleting: rotor type Type 45 Ti Pelleting: speed (g) 235000 Density gradient Type Discontinuous Number of initial discontinuous layers 6 Lowest density fraction 0% Highest density fraction 50% Total gradient volume, incl. sample (mL) 2.1 Sample volume (mL) 0.2 Orientation Bottom-up Speed (g) 120000 Duration (min) 1020 Fraction volume (mL) 0.2 Fraction processing Centrifugation Pelleting: volume per fraction 2.1 Pelleting: speed (g) 120000 Pelleting: adjusted k-factor TDB Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method Not determined Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis None

	EV230812	8/14	Streptomyces sp. Mg1	PSK0558	(d)(U)C DG Filtration	Hoefler BC	2017	57%
Study summary Full title A Link between Linearmycin Biosynthesis and Extracellular Vesicle Genesis Connects Specialized Metabolism and Bacterial Membrane Physiology. All authors Hoefler BC, Stubbendieck RM, Josyula NK, Moisan SM, Schulze EM, Straight PD Journal Cell Chem Biol Abstract Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and (show more...)Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and metal scavengers. Little is known about how specialized metabolites are processed and trafficked for their diverse competitive functions. Linearmycins A and B are linear polyketides with antifungal and antibacterial activity but are colony-localized in imaging mass spectrometry of Streptomyces sp. Mg1 (S. sp. Mg1). To decipher a connection between colony localization and antibiotic activity, we identified the linearmycin gene cluster and investigated linearmycin production and distribution by S. sp. Mg1. Our results uncover a large family of variant linearmycins with limited solubility in aqueous solution. We hypothesized that extracellular vesicles may traffic the lipid-like linearmycins. We found that vesicles isolated from culture supernatants contained linearmycins. Surprisingly, abolishing production of linearmycins in S. sp. Mg1 also diminished extracellular vesicle production. Our results reveal integration of linearmycin biosynthesis with production of extracellular vesicles, suggesting a deep connection between specialized metabolism and bacterial membrane physiology. (hide) EV-METRIC 57% (92nd percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin WT 96h Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Density gradient Filtration Protein markers EV: None non-EV: None Proteomics yes EV density (g/ml) not specified Show all info Study aim Biogenesis/cargo sorting/Identification of content (omics approaches) Sample Species Streptomyces sp. Mg1 Sample Type Cell culture supernatant EV-producing cells PSK0558 EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 10,000 g and 50,000 g Equal to or above 150,000 g Pelleting performed Yes Pelleting: rotor type Type 45 Ti Pelleting: speed (g) 235000 Density gradient Type Discontinuous Number of initial discontinuous layers 6 Lowest density fraction 0% Highest density fraction 50% Total gradient volume, incl. sample (mL) 2.1 Sample volume (mL) 0.2 Orientation Bottom-up Speed (g) 120000 Duration (min) 1020 Fraction volume (mL) 0.2 Fraction processing Centrifugation Pelleting: volume per fraction 2.1 Pelleting: speed (g) 120000 Pelleting: adjusted k-factor TDB Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method Not determined Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis None

	EV230812	9/14	Streptomyces sp. Mg1	PSK0558	(d)(U)C DG Filtration	Hoefler BC	2017	57%
Study summary Full title A Link between Linearmycin Biosynthesis and Extracellular Vesicle Genesis Connects Specialized Metabolism and Bacterial Membrane Physiology. All authors Hoefler BC, Stubbendieck RM, Josyula NK, Moisan SM, Schulze EM, Straight PD Journal Cell Chem Biol Abstract Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and (show more...)Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and metal scavengers. Little is known about how specialized metabolites are processed and trafficked for their diverse competitive functions. Linearmycins A and B are linear polyketides with antifungal and antibacterial activity but are colony-localized in imaging mass spectrometry of Streptomyces sp. Mg1 (S. sp. Mg1). To decipher a connection between colony localization and antibiotic activity, we identified the linearmycin gene cluster and investigated linearmycin production and distribution by S. sp. Mg1. Our results uncover a large family of variant linearmycins with limited solubility in aqueous solution. We hypothesized that extracellular vesicles may traffic the lipid-like linearmycins. We found that vesicles isolated from culture supernatants contained linearmycins. Surprisingly, abolishing production of linearmycins in S. sp. Mg1 also diminished extracellular vesicle production. Our results reveal integration of linearmycin biosynthesis with production of extracellular vesicles, suggesting a deep connection between specialized metabolism and bacterial membrane physiology. (hide) EV-METRIC 57% (92nd percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin ΔlnyI 24h Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Density gradient Filtration Protein markers EV: None non-EV: None Proteomics yes EV density (g/ml) not specified Show all info Study aim Biogenesis/cargo sorting/Identification of content (omics approaches) Sample Species Streptomyces sp. Mg1 Sample Type Cell culture supernatant EV-producing cells PSK0558 EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 10,000 g and 50,000 g Equal to or above 150,000 g Pelleting performed Yes Pelleting: rotor type Type 45 Ti Pelleting: speed (g) 235000 Density gradient Type Discontinuous Number of initial discontinuous layers 6 Lowest density fraction 0% Highest density fraction 50% Total gradient volume, incl. sample (mL) 2.1 Sample volume (mL) 0.2 Orientation Bottom-up Speed (g) 120000 Duration (min) 1020 Fraction volume (mL) 0.2 Fraction processing Centrifugation Pelleting: volume per fraction 2.1 Pelleting: speed (g) 120000 Pelleting: adjusted k-factor TDB Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method Not determined Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis None

	EV230812	10/14	Streptomyces sp. Mg1	PSK0558	(d)(U)C DG Filtration	Hoefler BC	2017	57%
Study summary Full title A Link between Linearmycin Biosynthesis and Extracellular Vesicle Genesis Connects Specialized Metabolism and Bacterial Membrane Physiology. All authors Hoefler BC, Stubbendieck RM, Josyula NK, Moisan SM, Schulze EM, Straight PD Journal Cell Chem Biol Abstract Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and (show more...)Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and metal scavengers. Little is known about how specialized metabolites are processed and trafficked for their diverse competitive functions. Linearmycins A and B are linear polyketides with antifungal and antibacterial activity but are colony-localized in imaging mass spectrometry of Streptomyces sp. Mg1 (S. sp. Mg1). To decipher a connection between colony localization and antibiotic activity, we identified the linearmycin gene cluster and investigated linearmycin production and distribution by S. sp. Mg1. Our results uncover a large family of variant linearmycins with limited solubility in aqueous solution. We hypothesized that extracellular vesicles may traffic the lipid-like linearmycins. We found that vesicles isolated from culture supernatants contained linearmycins. Surprisingly, abolishing production of linearmycins in S. sp. Mg1 also diminished extracellular vesicle production. Our results reveal integration of linearmycin biosynthesis with production of extracellular vesicles, suggesting a deep connection between specialized metabolism and bacterial membrane physiology. (hide) EV-METRIC 57% (92nd percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin ΔlnyI 36h Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Density gradient Filtration Protein markers EV: None non-EV: None Proteomics yes EV density (g/ml) not specified Show all info Study aim Biogenesis/cargo sorting/Identification of content (omics approaches) Sample Species Streptomyces sp. Mg1 Sample Type Cell culture supernatant EV-producing cells PSK0558 EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 10,000 g and 50,000 g Equal to or above 150,000 g Pelleting performed Yes Pelleting: rotor type Type 45 Ti Pelleting: speed (g) 235000 Density gradient Type Discontinuous Number of initial discontinuous layers 6 Lowest density fraction 0% Highest density fraction 50% Total gradient volume, incl. sample (mL) 2.1 Sample volume (mL) 0.2 Orientation Bottom-up Speed (g) 120000 Duration (min) 1020 Fraction volume (mL) 0.2 Fraction processing Centrifugation Pelleting: volume per fraction 2.1 Pelleting: speed (g) 120000 Pelleting: adjusted k-factor TDB Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method Not determined Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis None

	EV230812	11/14	Streptomyces sp. Mg1	PSK0558	(d)(U)C DG Filtration	Hoefler BC	2017	57%
Study summary Full title A Link between Linearmycin Biosynthesis and Extracellular Vesicle Genesis Connects Specialized Metabolism and Bacterial Membrane Physiology. All authors Hoefler BC, Stubbendieck RM, Josyula NK, Moisan SM, Schulze EM, Straight PD Journal Cell Chem Biol Abstract Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and (show more...)Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and metal scavengers. Little is known about how specialized metabolites are processed and trafficked for their diverse competitive functions. Linearmycins A and B are linear polyketides with antifungal and antibacterial activity but are colony-localized in imaging mass spectrometry of Streptomyces sp. Mg1 (S. sp. Mg1). To decipher a connection between colony localization and antibiotic activity, we identified the linearmycin gene cluster and investigated linearmycin production and distribution by S. sp. Mg1. Our results uncover a large family of variant linearmycins with limited solubility in aqueous solution. We hypothesized that extracellular vesicles may traffic the lipid-like linearmycins. We found that vesicles isolated from culture supernatants contained linearmycins. Surprisingly, abolishing production of linearmycins in S. sp. Mg1 also diminished extracellular vesicle production. Our results reveal integration of linearmycin biosynthesis with production of extracellular vesicles, suggesting a deep connection between specialized metabolism and bacterial membrane physiology. (hide) EV-METRIC 57% (92nd percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin ΔlnyI 48h Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Density gradient Filtration Protein markers EV: None non-EV: None Proteomics yes EV density (g/ml) not specified Show all info Study aim Biogenesis/cargo sorting/Identification of content (omics approaches) Sample Species Streptomyces sp. Mg1 Sample Type Cell culture supernatant EV-producing cells PSK0558 EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 10,000 g and 50,000 g Equal to or above 150,000 g Pelleting performed Yes Pelleting: rotor type Type 45 Ti Pelleting: speed (g) 235000 Density gradient Type Discontinuous Number of initial discontinuous layers 6 Lowest density fraction 0% Highest density fraction 50% Total gradient volume, incl. sample (mL) 2.1 Sample volume (mL) 0.2 Orientation Bottom-up Speed (g) 120000 Duration (min) 1020 Fraction volume (mL) 0.2 Fraction processing Centrifugation Pelleting: volume per fraction 2.1 Pelleting: speed (g) 120000 Pelleting: adjusted k-factor TDB Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method Not determined Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis None

	EV230812	12/14	Streptomyces sp. Mg1	PSK0558	(d)(U)C DG Filtration	Hoefler BC	2017	57%
Study summary Full title A Link between Linearmycin Biosynthesis and Extracellular Vesicle Genesis Connects Specialized Metabolism and Bacterial Membrane Physiology. All authors Hoefler BC, Stubbendieck RM, Josyula NK, Moisan SM, Schulze EM, Straight PD Journal Cell Chem Biol Abstract Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and (show more...)Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and metal scavengers. Little is known about how specialized metabolites are processed and trafficked for their diverse competitive functions. Linearmycins A and B are linear polyketides with antifungal and antibacterial activity but are colony-localized in imaging mass spectrometry of Streptomyces sp. Mg1 (S. sp. Mg1). To decipher a connection between colony localization and antibiotic activity, we identified the linearmycin gene cluster and investigated linearmycin production and distribution by S. sp. Mg1. Our results uncover a large family of variant linearmycins with limited solubility in aqueous solution. We hypothesized that extracellular vesicles may traffic the lipid-like linearmycins. We found that vesicles isolated from culture supernatants contained linearmycins. Surprisingly, abolishing production of linearmycins in S. sp. Mg1 also diminished extracellular vesicle production. Our results reveal integration of linearmycin biosynthesis with production of extracellular vesicles, suggesting a deep connection between specialized metabolism and bacterial membrane physiology. (hide) EV-METRIC 57% (92nd percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin ΔlnyI 72h Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Density gradient Filtration Protein markers EV: None non-EV: None Proteomics yes EV density (g/ml) not specified Show all info Study aim Biogenesis/cargo sorting/Identification of content (omics approaches) Sample Species Streptomyces sp. Mg1 Sample Type Cell culture supernatant EV-producing cells PSK0558 EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 10,000 g and 50,000 g Equal to or above 150,000 g Pelleting performed Yes Pelleting: rotor type Type 45 Ti Pelleting: speed (g) 235000 Density gradient Type Discontinuous Number of initial discontinuous layers 6 Lowest density fraction 0% Highest density fraction 50% Total gradient volume, incl. sample (mL) 2.1 Sample volume (mL) 0.2 Orientation Bottom-up Speed (g) 120000 Duration (min) 1020 Fraction volume (mL) 0.2 Fraction processing Centrifugation Pelleting: volume per fraction 2.1 Pelleting: speed (g) 120000 Pelleting: adjusted k-factor TDB Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method Not determined Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis None

	EV230812	13/14	Streptomyces sp. Mg1	PSK0558	(d)(U)C DG Filtration	Hoefler BC	2017	57%
Study summary Full title A Link between Linearmycin Biosynthesis and Extracellular Vesicle Genesis Connects Specialized Metabolism and Bacterial Membrane Physiology. All authors Hoefler BC, Stubbendieck RM, Josyula NK, Moisan SM, Schulze EM, Straight PD Journal Cell Chem Biol Abstract Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and (show more...)Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and metal scavengers. Little is known about how specialized metabolites are processed and trafficked for their diverse competitive functions. Linearmycins A and B are linear polyketides with antifungal and antibacterial activity but are colony-localized in imaging mass spectrometry of Streptomyces sp. Mg1 (S. sp. Mg1). To decipher a connection between colony localization and antibiotic activity, we identified the linearmycin gene cluster and investigated linearmycin production and distribution by S. sp. Mg1. Our results uncover a large family of variant linearmycins with limited solubility in aqueous solution. We hypothesized that extracellular vesicles may traffic the lipid-like linearmycins. We found that vesicles isolated from culture supernatants contained linearmycins. Surprisingly, abolishing production of linearmycins in S. sp. Mg1 also diminished extracellular vesicle production. Our results reveal integration of linearmycin biosynthesis with production of extracellular vesicles, suggesting a deep connection between specialized metabolism and bacterial membrane physiology. (hide) EV-METRIC 57% (92nd percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin ΔlnyI 96h Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Density gradient Filtration Protein markers EV: None non-EV: None Proteomics yes EV density (g/ml) not specified Show all info Study aim Biogenesis/cargo sorting/Identification of content (omics approaches) Sample Species Streptomyces sp. Mg1 Sample Type Cell culture supernatant EV-producing cells PSK0558 EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 10,000 g and 50,000 g Equal to or above 150,000 g Pelleting performed Yes Pelleting: rotor type Type 45 Ti Pelleting: speed (g) 235000 Density gradient Type Discontinuous Number of initial discontinuous layers 6 Lowest density fraction 0% Highest density fraction 50% Total gradient volume, incl. sample (mL) 2.1 Sample volume (mL) 0.2 Orientation Bottom-up Speed (g) 120000 Duration (min) 1020 Fraction volume (mL) 0.2 Fraction processing Centrifugation Pelleting: volume per fraction 2.1 Pelleting: speed (g) 120000 Pelleting: adjusted k-factor TDB Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method Not determined Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis None

	EV230812	14/14	Streptomyces aizunensis	PDS0116	(d)(U)C DG Filtration	Hoefler BC	2017	57%
Study summary Full title A Link between Linearmycin Biosynthesis and Extracellular Vesicle Genesis Connects Specialized Metabolism and Bacterial Membrane Physiology. All authors Hoefler BC, Stubbendieck RM, Josyula NK, Moisan SM, Schulze EM, Straight PD Journal Cell Chem Biol Abstract Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and (show more...)Specialized metabolites support bacterial competitive fitness as antibiotics, signals, pigments, and metal scavengers. Little is known about how specialized metabolites are processed and trafficked for their diverse competitive functions. Linearmycins A and B are linear polyketides with antifungal and antibacterial activity but are colony-localized in imaging mass spectrometry of Streptomyces sp. Mg1 (S. sp. Mg1). To decipher a connection between colony localization and antibiotic activity, we identified the linearmycin gene cluster and investigated linearmycin production and distribution by S. sp. Mg1. Our results uncover a large family of variant linearmycins with limited solubility in aqueous solution. We hypothesized that extracellular vesicles may traffic the lipid-like linearmycins. We found that vesicles isolated from culture supernatants contained linearmycins. Surprisingly, abolishing production of linearmycins in S. sp. Mg1 also diminished extracellular vesicle production. Our results reveal integration of linearmycin biosynthesis with production of extracellular vesicles, suggesting a deep connection between specialized metabolism and bacterial membrane physiology. (hide) EV-METRIC 57% (92nd percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin Control condition Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Density gradient Filtration Protein markers EV: None non-EV: None Proteomics yes EV density (g/ml) not specified Show all info Study aim Biogenesis/cargo sorting/Identification of content (omics approaches) Sample Species Streptomyces aizunensis Sample Type Cell culture supernatant EV-producing cells PDS0116 EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 10,000 g and 50,000 g Equal to or above 150,000 g Pelleting performed Yes Pelleting: rotor type Type 45 Ti Pelleting: speed (g) 235000 Density gradient Type Discontinuous Number of initial discontinuous layers 8 Lowest density fraction 10% Highest density fraction 50% Total gradient volume, incl. sample (mL) 14 Sample volume (mL) 1 Orientation Bottom-up Speed (g) 100400 Duration (min) 960 Fraction volume (mL) 1 Fraction processing Centrifugation Pelleting: volume per fraction 4mL Pelleting: speed (g) 120000 Pelleting: adjusted k-factor 1.857 Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method Not determined Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis None

				1 - 14 of 14

this entry contains some non-printable text. Please contact the admin. 'ascii' codec can't encode character '\u0394' in position 203: ordinal not in range(128)

EV-TRACK ID	EV230812
species	Streptomyces sp. Mg1	Streptomyces sp. Mg1	Streptomyces sp. Mg1	Streptomyces sp. Mg1	Streptomyces sp. Mg1	Streptomyces sp. Mg1	Streptomyces sp. Mg1	Streptomyces sp. Mg1	Streptomyces sp. Mg1	Streptomyces sp. Mg1	Streptomyces sp. Mg1	Streptomyces sp. Mg1	Streptomyces sp. Mg1	Streptomyces aizunensis
sample type	Cell culture	Cell culture	Cell culture	Cell culture	Cell culture	Cell culture	Cell culture	Cell culture	Cell culture	Cell culture	Cell culture	Cell culture	Cell culture	Cell culture
cell type	PSK0558	PSK0558	PSK0558	PSK0558	PSK0558	PSK0558	PSK0558	PSK0558	PSK0558	PSK0558	PSK0558	PSK0558	PSK0558	PDS0116

Study summary

Study data

Study summary

Study data

Study summary

Study data

Study summary

Study data

Study summary

Study data

Study summary

Study data

Study summary

Study data

Study summary

Study data

Study summary

Study data

Study summary

Study data

Study summary

Study data

Study summary

Study data

Study summary

Study data

Study summary

Study data