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The immune function of MHC class II molecules mutated in the putative superdimer interface

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Abstract

Analysis of the crystal structure of human class II (HLA-DR1) molecules suggests that the αβ heterodimer may be further ordered as a dimer of heterodimers (superdimer), leading to the hypothesis that T cell receptor dimerisation is a mechanism for initiating signaling events preceding T cell activation. The interface between pairs of molecules is stabilised by both salt bridges, polar and hydrophobic interactions. The residues that form the superdimer interface occur in three areas distinct from the antigen-binding groove. They can be defined as follows: region 1, β-β contacts in the helix of the β1 domain; region 2, α-α contacts near the α1/α2 domain junction and region 3; α-β contacts in the α2/β2 domains adjacent to the plasma membrane. To determine whether salt bridges and polar interactions formed within these regions are involved in the immune function of the murine MHC class II molecule, I-Ab, appropriate residues in both the α and β chain were identified and mutated to uncharged alanine. Cell lines transfected with different combinations of mutated α and β chains were generated and tested for MHC class II expression, peptide binding capabilities, and ability to present antigenic peptide to an OVA-specific T cell hybridoma. With the exception of two residues in region 2, the substitutions tested did not modulate MHC class II expression, or peptide binding function. When tested for ability to present peptide to an antigen-specific T cell hybridoma, with the exception of mutations in region 2, the substitutions did not appear to abrogate the ability of I-Ab to stimulate the T cells. These results suggest that mutation of residues in region 2 of the putative superdimer interface have a gross effect on the ability of I-Ab to be expressed on the cell surface. However, abrogation of salt bridges in region 1 and 3 do not influence I-Ab cell surface expression, peptide binding or ability to stimulate antigen-specific T cells.

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Authors and Affiliations

  1. School of Pharmacy and Medical Sciences, University of South Australia, North Tce, Adelaide, South Australia, Australia

    John D. Hayball

  2. Dame Roma Mitchell Cancer Research Laboratories, Hanson Institute, Frome Road, Adelaide, South Australia, Australia

    John D. Hayball

  3. Queen Elizabeth II Medical School, University Department of Medicine, Nedlands, Perth, Western Australia, Australia

    Richard A. Lake

  4. School of Pharmacy and Medical Sciences, University of South Australia, GPO Box 2471, Adelaide, South Australia, 5001, Australia

    John D. Hayball

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Hayball, J.D., Lake, R.A. The immune function of MHC class II molecules mutated in the putative superdimer interface. Mol Cell Biochem 273, 1–9 (2005). https://doi.org/10.1007/s11010-005-5281-4

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  • DOI: https://doi.org/10.1007/s11010-005-5281-4

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