Studying ubiquitination of MHC class I molecules

Methods Mol Biol. 2013:960:109-125. doi: 10.1007/978-1-62703-218-6_9.

Abstract

The covalent attachment of ubiquitin to a protein is one of the most common post-translational modifications and regulates diverse eukaryotic cellular processes. Ubiquitination of MHC class I was first described in the context of viral proteins which target MHC class I for degradation in the endoplasmic reticulum and at the cell surface. Study of viral-induced MHC class I degradation has been extremely instructive in elucidating cellular pathways for degradation of membrane and secretory proteins. More recently, ubiquitination of endogenous MHC class I heavy chains which fail to achieve their native conformation and undergo endoplasmic-reticulum associated degradation has been demonstrated.In this chapter we describe methods for identification of endogenous ubiquitinated MHC class I heavy chains by MHC class I-immunoprecipitation and ubiquitin-specific immunoblot or by metabolic labeling and immunoprecipitation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Separation
  • Electrophoresis, Polyacrylamide Gel
  • HeLa Cells
  • Histocompatibility Antigens Class I / chemistry
  • Histocompatibility Antigens Class I / metabolism*
  • Humans
  • Immunoblotting
  • Immunoprecipitation
  • Membranes, Artificial
  • Protein Denaturation
  • Ubiquitination*
  • Viral Proteins / metabolism

Substances

  • Histocompatibility Antigens Class I
  • Membranes, Artificial
  • Viral Proteins