title: Studying MHC class I peptide loading and exchange in vitro.
abstract: In the endoplasmic reticulum (ER), MHC class I molecules associate with several specialized proteins, forming a large macromolecular complex referred to as the "peptide-loading complex" (PLC). In the PLC, antigenic peptides undergo a stringent selection process for binding onto MHC class I molecules. This ensures that the immune system elicits robust CD8+ T-cell responses to viruses and solid tumors. The ability to reconstitute in vitro MHC class I molecules in association with key proteins of the PLC provides a mean for studying at the molecular level how antigenic peptides are selected for presentation to CD8+ T-cells. Here, we describe practical procedures for generating a cell-free system involving MHC class I molecules and tapasin, a critical protein of the PLC, that can be used as a versatile tool for biochemical and mechanistic studies of peptide loading and exchange.
authors: Department of Microbiology and Immunology, University of Illinois at Chicago, College of Medicine, Chicago, IL, USA. mbouvier@uic.edu.
authors: Marlene
authors: M
authors: Bouvier
mesh_terms: D017951:Antigen Presentation; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002474:Cell-Free System / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D015395:Histocompatibility Antigens Class I / Q000276:immunology / Q000378:metabolism*; D007111:Immunity, Cellular; D002479:Inclusion Bodies / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D055550:Protein Stability; D012995:Solubility
chemical_list: D015395:Histocompatibility Antigens Class I; D026901:Membrane Transport Proteins; D010455:Peptides; C107762:tapasin