Role of the rab5 guanine nucleotide exchange factor, Rme-6, in the regulation of clathrin-coated vesicle uncoating

Methods Mol Biol. 2015:1298:283-93. doi: 10.1007/978-1-4939-2569-8_24.

Abstract

Clathrin-coated pits are major ports of entry into eukaryotic cells. Following scission of clathrin-coated pits to form coated vesicles, the peripheral membrane proteins that form the coat need to be removed. Here we describe an assay that provides a measure of the extent of clathrin coat uncoating in intact cells. This assay has been used to investigate rab5-dependent modulation of uncoating. Specifically, it has been used to identify functional differences between rab5 guanine nucleotide exchange factors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biological Assay / methods*
  • Cell Culture Techniques
  • Clathrin-Coated Vesicles / metabolism*
  • Fluorescent Antibody Technique
  • Gene Knockdown Techniques
  • Guanine Nucleotide Exchange Factors / deficiency
  • Guanine Nucleotide Exchange Factors / genetics
  • Guanine Nucleotide Exchange Factors / metabolism*
  • HEK293 Cells
  • Humans
  • Image Processing, Computer-Assisted
  • RNA, Small Interfering / genetics
  • Transfection

Substances

  • GAPVD1 protein, human
  • Guanine Nucleotide Exchange Factors
  • RNA, Small Interfering