title: Studying ubiquitination of MHC class I molecules.
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.
authors: Cambridge Institute for Medical Research, University of Cambridge, Addenbrooke's Hospital, Cambridge, UK.
authors: Marian L
authors: ML
authors: Burr
authors: Cambridge Institute for Medical Research, University of Cambridge, Addenbrooke's Hospital, Cambridge, UK.
authors: Jessica M
authors: JM
authors: Boname
authors: Cambridge Institute for Medical Research, Addenbrooke's Hospital, University of Cambridge, Cambridge, UK. pjl30@cam.ac.uk.
authors: Paul J
authors: PJ
authors: Lehner
mesh_terms: D002469:Cell Separation; D004591:Electrophoresis, Polyacrylamide Gel; D006367:HeLa Cells; D015395:Histocompatibility Antigens Class I / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D008567:Membranes, Artificial; D011489:Protein Denaturation; D054875:Ubiquitination; D014764:Viral Proteins / Q000378:metabolism
chemical_list: D015395:Histocompatibility Antigens Class I; D008567:Membranes, Artificial; D014764:Viral Proteins