Proteomics

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Heart extracellular vesicle proteome

ABSTRACT: Cardiac function is dynamically regulated by intercellular signalling. As an important intercellular signalling mediator, this study focused on isolating and defining membrane bound signalling packages termed extracellular vesicles (EVs) directly from heart. Here, we utilised a gentle, enzymatic perfusion to extract EVs from whole cardiac tissues (cEVs) for biophysical and proteomic characterisation; cEVs (100-300 nm) are Cd63+, Tsg101+, Pdcd6ip/Alix+ (EV markers) and contain proteins associated with cardiomyocyte (Actn1, Myh6), cardiac progenitor (Cd34, Abcg2), smooth muscle (Cald1, Tagln), endothelial (Vwf), and fibroblast (Ckap4) origins. Proteomic profiling of cEVs revealed 1721 proteins, which bioinformatics revealed hold an enrichment in metabolic functions of glycolysis, fatty acid oxidation, and antioxidant action, presumably to meet the high energetic demands and balance resulting oxidative stress in cardiac tissue.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Heart

DISEASE(S): Disease Free

SUBMITTER: david greening  

LAB HEAD: David Greening

PROVIDER: PXD023570 | Pride | 2021-04-26

REPOSITORIES: Pride

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Publications

Proteome characterisation of extracellular vesicles isolated from heart.

Claridge Bethany B   Rai Alin A   Fang Haoyun H   Matsumoto Aya A   Luo Jieting J   McMullen Julie R JR   Greening David W DW  

Proteomics 20210506 13-14

Cardiac intercellular communication is critical for heart function and often dysregulated in cardiovascular diseases. While cardiac extracellular vesicles (cEVs) are emerging mediators of signalling, their isolation remains a technical challenge hindering our understanding of cEV protein composition. Here, we utilised Langendorff-collagenase-based enzymatic perfusion and differential centrifugation to isolate cEVs from mouse heart (yield 3-6 μg/heart). cEVs are ∼200 nm, express classical EV mark  ...[more]

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