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M94A0085.TXT
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1994-10-01
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Document 0085
DOCN M94A0085
TI Mutation of the alpha 2 domain disulfide bridge of the class I molecule
HLA-A*0201. Effect on maturation and peptide presentation.
DT 9412
AU Warburton RJ; Matsui M; Rowland-Jones SL; Gammon MC; Katzenstein GE; Wei
T; Edidin M; Zweerink HJ; McMichael AJ; Frelinger JA; Department of
Microbiology and Immunology, University of North; Carolina, Chapel Hill
27599-7290.
SO Hum Immunol. 1994 Apr;39(4):261-71. Unique Identifier : AIDSLINE
MED/94350693
AB A combination of saturation and site-directed mutagenesis was utilized
to disrupt the alpha 2 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.
DE Amino Acid Sequence Antigen Presentation/*IMMUNOLOGY Blotting, Western
Cell Line Cells, Cultured Cytotoxicity, Immunologic Disulfides Gene
Products, pol/CHEMICAL SYNTHESIS/*IMMUNOLOGY Human HIV-1/IMMUNOLOGY
HLA-A Antigens/GENETICS/*IMMUNOLOGY Molecular Sequence Data
Mutagenesis, Site-Directed *Mutation Oligopeptides/CHEMICAL
SYNTHESIS/*IMMUNOLOGY Support, Non-U.S. Gov't Support, U.S. Gov't,
P.H.S. T-Lymphocytes, Cytotoxic/IMMUNOLOGY Viral Matrix
Proteins/CHEMICAL SYNTHESIS/*IMMUNOLOGY JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).