In Vitro Studies of MHC Class I Peptide Loading and Exchange.
Fluorescence anisotropy
MHC class I molecule
Peptide association
Peptide dissociation
Peptide exchange
Peptide-loading complex
Tapasin
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2019
2019
Historique:
entrez:
1
6
2019
pubmed:
31
5
2019
medline:
23
1
2020
Statut:
ppublish
Résumé
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 that determines which antigen becomes part of the repertoire presented by 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 made up of MHC class I molecules and tapasin that can be used for mechanistic studies of peptide loading and exchange.
Identifiants
pubmed: 31147933
doi: 10.1007/978-1-4939-9450-2_6
pmc: PMC7278038
mid: NIHMS1594059
doi:
Substances chimiques
Histocompatibility Antigens Class I
0
Membrane Transport Proteins
0
Peptides
0
Proto-Oncogene Proteins c-fos
0
beta 2-Microglobulin
0
tapasin
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
71-81Subventions
Organisme : NIAID NIH HHS
ID : R01 AI045070
Pays : United States
Organisme : NIAID NIH HHS
ID : R01 AI108546
Pays : United States
Organisme : NIAID NIH HHS
ID : R01 AI114467
Pays : United States
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