Abstract
Presentation of antigenic peptides to CTLs at the cell surface first requires assembly of MHC class I with peptide and beta 2-microglobulin in the endoplasmic reticulum. This process involves an assembly complex of several proteins, including TAP, tapasin, and calreticulin, all of which associate specifically with the beta 2-microglobulin-assembled, open form of the class I heavy chain. To better comprehend at a molecular level the regulation of class I assembly, we have assessed the influence of multiple individual amino acid substitutions in the MHC class I alpha 2 domain on interaction with TAP, tapasin, and calreticulin. In this report, we present evidence indicating that many residues surrounding position 134 in H-2Ld influence interaction with assembly complex components. Most mutations decreased association, but one (LdK131D) strongly increased it. The Ld mutants, with the exception of LdK131D, exhibited characteristics suggesting suboptimal intracellular peptide loading, similar to the phenotype of Ld expressed in a tapasin-deficient cell line. Notably, K131D was less peptide inducible than wild-type Ld, which is consistent with its unusually strong association with the endoplasmic reticulum assembly complex.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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ATP Binding Cassette Transporter, Subfamily B, Member 2
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ATP-Binding Cassette Transporters / metabolism
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Amino Acid Sequence
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Amino Acid Substitution / genetics
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Amino Acid Substitution / immunology
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Animals
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Antigen Presentation*
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Aspartic Acid / genetics
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Calcium-Binding Proteins / metabolism
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Calreticulin
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Cell Line, Transformed
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H-2 Antigens / chemistry
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H-2 Antigens / genetics
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H-2 Antigens / metabolism
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HLA Antigens / chemistry
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HLA Antigens / genetics
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HLA Antigens / metabolism
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Histocompatibility Antigen H-2D
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Histocompatibility Antigens Class I / chemistry*
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Histocompatibility Antigens Class I / genetics
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Histocompatibility Antigens Class I / metabolism
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Humans
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Lysine / genetics
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Membrane Proteins / genetics
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Membrane Proteins / immunology
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Membrane Proteins / metabolism
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Mice
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Molecular Chaperones / metabolism
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Molecular Sequence Data
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Mutagenesis, Site-Directed
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Peptide Fragments / chemistry
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Peptide Fragments / genetics
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Peptide Fragments / immunology*
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Peptide Fragments / metabolism
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Protein Folding
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Ribonucleoproteins / metabolism
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beta 2-Microglobulin / metabolism
Substances
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ATP Binding Cassette Transporter, Subfamily B, Member 2
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ATP-Binding Cassette Transporters
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Calcium-Binding Proteins
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Calreticulin
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H-2 Antigens
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HLA Antigens
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Histocompatibility Antigen H-2D
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Histocompatibility Antigens Class I
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Membrane Proteins
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Molecular Chaperones
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Peptide Fragments
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Ribonucleoproteins
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TAP1 protein, human
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Tap1 protein, mouse
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beta 2-Microglobulin
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Aspartic Acid
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Lysine