Measuring synthesis and degradation of MHC class I molecules

Methods Mol Biol. 2013:960:93-108. doi: 10.1007/978-1-62703-218-6_8.

Abstract

Major histocompatibility complex (MHC) class I molecules function to present pathogen-derived peptides to cytotoxic T cells or act as ligands for Natural Killer cells, thus alerting the immune system to the presence of invading pathogens. Furthermore MHC class I molecules can be strongly associated with autoimmune diseases. Therefore understanding not only the biosynthesis and the degradation pathways of MHC class I molecules has become important in determining their role in pathogen and autoimmune-related diseases. Here we describe how using epitope-tagged MHC class I molecules can aid in the analysis of MHC class I molecule biosynthesis and degradation and also complement studies using conventional conformationally specific antibodies. Coupled together with pharmacological manipulation which can target both biosynthetic and degradative pathways, this offers a powerful tool in analyzing MHC class I molecules.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Electrophoresis, Polyacrylamide Gel
  • Epitopes / immunology
  • Flow Cytometry
  • HEK293 Cells
  • HeLa Cells
  • Histocompatibility Antigens Class I / biosynthesis*
  • Histocompatibility Antigens Class I / genetics
  • Histocompatibility Antigens Class I / immunology
  • Histocompatibility Antigens Class I / metabolism*
  • Humans
  • Immunoblotting
  • Immunoprecipitation
  • Proteolysis* / drug effects

Substances

  • Epitopes
  • Histocompatibility Antigens Class I