TY - JOUR
T1 - Mutation of the α2 domain disulfide bridge of the class I molecule HLA-A*0201 Effect on maturation and peptide presentation
AU - Warburton, Robert J.
AU - Matsui, Masanori
AU - Rowland-Jones, Sarah L.
AU - Gammon, Maureen C.
AU - Katzenstein, Gil E.
AU - Wei, Taiyin
AU - Edidin, Michael
AU - Zweerink, Hans J.
AU - McMichael, Andrew J.
AU - Frelinger, Jeffrey A.
N1 - Funding Information:
was supported by grants AI-20288 and AI-29324 from the National Institute of Allergy and Infectious Diseases, the Medical Research Council, and the North Atlantic Treaty Or-gamzation
Copyright:
Copyright 2014 Elsevier B.V., All rights reserved.
PY - 1994/4
Y1 - 1994/4
N2 - A combination of saturation and site-directed mutagenesis was utilized to disrupt the α2 domain disulfide bridge of HLA-A*0201. Mutation of cysteine 101 to a serine (C101S) or of cysteine 164 to alanine (C164A) decreased the rate of maturation of the heavy chain, the total amount of mature heavy chain within the cell, and the level of surface expression. Cells expressing these genes and loaded with a synthetic peptide derived from the influenza A matrix protein (58-66) were recognized poorly by HLA-A*0201-restricted, peptide-specific CTLs. Cells expressing mutant HLA-A*0201 loaded with a synthetic peptide derived from the HIV-1 pol protein (476-484) were not recognized by pol IV-9-specific CTLs. Mutant C164A cells infected with influenza virus were partially recognized by influenza matrix peptide-specific CTLs, while C101S cells were not lysed. Surprisingly, endogenous peptide loading of cells expressing mutant HLA-A*0201 using a minigene coding for either the influenza A matrix peptide 58-66, or HIV-1 pol peptide 476-484, resulted in efficient CTL recognition. This suggests different structural constraints for peptide binding in the endoplasmic reticulum during biosynthesis and for binding to exported molecules on the cells surface.
AB - A combination of saturation and site-directed mutagenesis was utilized to disrupt the α2 domain disulfide bridge of HLA-A*0201. Mutation of cysteine 101 to a serine (C101S) or of cysteine 164 to alanine (C164A) decreased the rate of maturation of the heavy chain, the total amount of mature heavy chain within the cell, and the level of surface expression. Cells expressing these genes and loaded with a synthetic peptide derived from the influenza A matrix protein (58-66) were recognized poorly by HLA-A*0201-restricted, peptide-specific CTLs. Cells expressing mutant HLA-A*0201 loaded with a synthetic peptide derived from the HIV-1 pol protein (476-484) were not recognized by pol IV-9-specific CTLs. Mutant C164A cells infected with influenza virus were partially recognized by influenza matrix peptide-specific CTLs, while C101S cells were not lysed. Surprisingly, endogenous peptide loading of cells expressing mutant HLA-A*0201 using a minigene coding for either the influenza A matrix peptide 58-66, or HIV-1 pol peptide 476-484, resulted in efficient CTL recognition. This suggests different structural constraints for peptide binding in the endoplasmic reticulum during biosynthesis and for binding to exported molecules on the cells surface.
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U2 - 10.1016/0198-8859(94)90269-0
DO - 10.1016/0198-8859(94)90269-0
M3 - Article
C2 - 8071101
AN - SCOPUS:0028177703
SN - 0198-8859
VL - 39
SP - 261
EP - 271
JO - Human Immunology
JF - Human Immunology
IS - 4
ER -