Cutting edge: HLA-DM functions through a mechanism that does not require specific conserved hydrogen bonds in class II MHC-peptide complexes.
HLA-DM catalyzes peptide dissociation and exchange in class II MHC molecules through a mechanism that has been proposed to involve the disruption of specific components of the conserved hydrogen bond network in MHC-peptide complexes. HLA-DR1 molecules with alanine substitutions at each of the six conserved H- bonding positions were expressed in cells, and susceptibility to DM catalytic activity was evaluated by measuring the release of CLIP. The mutants alphaN62A, alphaN69A, alphaR76A, and betaH81A DR1 were fully susceptible to DM-mediated CLIP release, and betaN82A resulted in spontaneous release of CLIP. Using recombinant soluble DR1 molecules, the amino acid betaN82 was observed to contribute disproportionately in stabilizing peptide complexes. Remarkably, the catalytic potency of DM with each beta-chain mutant was equal to or greater than that observed with wild-type DR1. Our results support the conclusion that no individual component of the conserved hydrogen bond network plays an essential role in the DM catalytic mechanism.
Zhou
Zemin
Z
Department of Pathology, University of Utah, Salt Lake City, UT 84112, USA.
Callaway
Kari A
KA
Weber
Dominique A
DA
Jensen
Peter E
PE
D019943:Amino Acid Substitution / Q000235:genetics; D000944:Antigens, Differentiation, B-Lymphocyte / Q000378:metabolism; D002384:Catalysis; D017124:Conserved Sequence / Q000235:genetics / Q000276:immunology*; D006681:HLA-D Antigens / Q000737:chemistry* / Q000235:genetics* / Q000502:physiology; D006684:HLA-DR Antigens / Q000235:genetics / Q000378:metabolism; D015798:HLA-DR1 Antigen / Q000235:genetics / Q000378:metabolism; D059811:HLA-DRB1 Chains; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000378:metabolism; D000949:Histocompatibility Antigens Class II / Q000378:metabolism; D006801:Humans; D006860:Hydrogen Bonding; D021122:Protein Subunits / Q000378:metabolism
D000944:Antigens, Differentiation, B-Lymphocyte; D006681:HLA-D Antigens; C084245:HLA-DM antigens; D006684:HLA-DR Antigens; D015798:HLA-DR1 Antigen; D059811:HLA-DRB1 Chains; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D000949:Histocompatibility Antigens Class II; D021122:Protein Subunits; C045084:invariant chain