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).