Abstract
The peptide editor HLA-DM (DM) mediates exchange of peptides bound to major histocompatibility (MHC) class II molecules during antigen processing; however, the mechanism by which DM displaces peptides remains unclear. Here we generated a soluble mutant HLA-DR1 with a histidine-to-asparagine substitution at position 81 of the β-chain (DR1βH81N) to perturb an important hydrogen bond between MHC class II and peptide. Peptide–DR1βH81N complexes dissociated at rates similar to the dissociation rates of DM-induced peptide–wild-type DR1, and DM did not enhance the dissociation of peptide–DR1βH81N complexes. Reintroduction of an appropriate hydrogen bond (DR1βH81N βV85H) restored DM-mediated peptide dissociation. Thus, DR1βH81N might represent a 'post-DM effect' conformation. We suggest that DM may mediate peptide dissociation by a 'hit-and-run' mechanism that results in conformational changes in MHC class II molecules and disruption of hydrogen bonds between βHis81 and bound peptide.
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Acknowledgements
We thank L.J. Stern and E.D. Mellins for discussions; K. Su for help with the experimental setup, C.-H. (Bear) Huang for help with modeling; and Erika Darrah for critical reading of the manuscript. P. Roche (National Cancer Institute) donated cDNA constructs for the HLA-DM α-and β-chains. K.N. dedicates dedicate this work to the memory of Varun Shankar, who passed away during the preparation of this manuscript. Supported by the National Institutes of Health (R01AI063764 and R01GM53549 to S.S.-N.).
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Supplementary information
Supplementary Fig. 1
Size separation profiles of double mutant DR1 proteins are identical to DR1WT and DR1βH81N. (PDF 306 kb)
Supplementary Fig. 2
SDS stability of the double mutant DR1 proteins. (PDF 274 kb)
Supplementary Fig. 3
Both DR1(βH81N βV85H) and DR1(βH81N βN82H) dissociate from HA(anchorless) rapidly in an SPR experiment. (PDF 152 kb)
Supplementary Fig. 4
Model of DM-mediated peptide dissociation and epitope selection in the antigen-presenting cell. (PDF 230 kb)
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Narayan, K., Chou, CL., Kim, A. et al. HLA-DM targets the hydrogen bond between the histidine at position β81 and peptide to dissociate HLA-DR–peptide complexes. Nat Immunol 8, 92–100 (2007). https://doi.org/10.1038/ni1414
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DOI: https://doi.org/10.1038/ni1414
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