Identification of Podocyte Cargo Proteins by Proteomic Analysis of Clathrin-Coated Vesicles.

ABSTRACT:

Background

Clathrin-mediated endocytosis (CME) plays a fundamental role in podocyte health. Genetic ablation of genes implicated in CME has been shown to cause severe proteinuria and foot process effacement in mice. However, little is known about the cargo of clathrin-coated vesicles (CCVs) in podocytes. The goal of this study was to isolate CCVs from podocytes and identify their cargo by proteomic analysis.

Methods

Glomeruli isolated from Podocin-Cre Rosa-DTR^flox mouse kidneys were seeded and treated with diphtheria toxin to obtain pure primary podocyte cultures. CCVs were isolated by differential gradient ultracentrifugation, and enrichment of CCVs was assessed by immunoblotting and electron microscopy (EM). Liquid chromatography-mass spectrometry

SUBMITTER: Groener M 

PROVIDER: S-EPMC8809311 | biostudies-literature | 2020 Jun

REPOSITORIES: biostudies-literature