Proteomics

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Surface protein profiling of prostate derived extracellular vesicles


ABSTRACT: Extracellular vesicles (EVs) are mediators of intercellular communication and a promising class of biomarkers. Surface proteins of EV play decisive roles in establishing a connection with recipient cells, and they are putative targets in diagnostic assays. Analysis of the surface proteins can thus both illuminate the biological functions of EVs and help identify potential biomarkers. The surface proteins of intact seminal fluid ssmall EV (SF-sEVs and PC3) sEVs were labeled using sulfo-NHS-SS-biotin, a membrane-impermeable reagent reacting with primary amines. The fractions obtained from the purification process were: (i) protein in total sEV lysate (Total); (ii) proteins isolated from sEV surfaces (Surface), and (iii) supernatant proteins left after isolation of surface proteins (Total minus Surface). Proteins from each fraction were digested by trypsin and analyzed by HRMS. A total PC3 cell lysate was prepared andanalyzed as a control.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Claudia Fredolini  

LAB HEAD: Masood Kamali-Moghaddam

PROVIDER: PXD037791 | Pride | 2023-03-11

REPOSITORIES: Pride

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Publications


Extracellular vesicles (EVs) are mediators of intercellular communication and a promising class of biomarkers. Surface proteins of EVs play decisive roles in establishing a connection with recipient cells, and they are putative targets for diagnostic assays. Analysis of the surface proteins can thus both illuminate the biological functions of EVs and help identify potential biomarkers. We developed a strategy combining high-resolution mass spectrometry (HRMS) and  proximity ligation assays (PLA)  ...[more]

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