A triad of molecular regions contribute to the formation of two distinct MHC class II conformers

Mol Immunol. 2016 Jun;74:59-70. doi: 10.1016/j.molimm.2016.04.010. Epub 2016 May 2.

Abstract

MHC class II molecules present antigen-derived peptides to CD4T cells to drive the adaptive immune response. Previous work has established that class II αβ dimers can adopt two distinct conformations, driven by the differential pairing of transmembrane domain GxxxG dimerization motifs. These class II conformers differ in their ability to be loaded with antigen-derived peptide and to effectively engage CD4T cells. Motif 1 (M1) paired I-A(k) class II molecules are efficiently loaded with peptides derived from the processing of B cell receptor-bound antigen, have unique B cell signaling properties and high T cell stimulation activity. The 11-5.2mAb selectively binds M1 paired I-A(k) class II molecules. However, the molecular determinants of 11-5.2 binding are currently unclear. Here, we report the ability of a human class II transmembrane domain to drive both M1 and M2 class II conformer formation. Protease sensitivity analysis further strengthens the idea that there are conformational differences between the extracellular domains of M1 and M2 paired class II. Finally, MHC class II chain alignments and site directed mutagenesis reveals a triad of molecular regions that contributes to 11-5.2mAb binding. In addition to transmembrane GxxxG motif domain pairing, 11-5.2 binding is influenced directly by α chain residue Glu-71 and indirectly by the region around the inter-chain salt bridge formed by α chain Arg-52 and β chain Glu-86. These findings provide insight into the complexity of 11-5.2mAb recognition of the M1 paired I-A(k) class II conformer and further highlight the molecular heterogeneity of peptide-MHC class II complexes that drive T cell antigen recognition.

Keywords: Antigen presentation; Conformation; Epitope; GxxxG motif; MHC class II.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antibodies, Monoclonal
  • Antigen Presentation / immunology
  • Blotting, Western
  • CD4-Positive T-Lymphocytes / immunology
  • Histocompatibility Antigens Class II / chemistry*
  • Histocompatibility Antigens Class II / immunology*
  • Humans
  • Lymphocyte Activation / immunology
  • Mice
  • Mice, Inbred BALB C
  • Molecular Conformation
  • Mutagenesis, Site-Directed

Substances

  • Antibodies, Monoclonal
  • Histocompatibility Antigens Class II