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HLA-DM targets the hydrogen bond between the histidine at position β81 and peptide to dissociate HLA-DR–peptide complexes

Nature Immunology volume 8, pages 92–100 (2007)Cite this article

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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Figure 1: DR1βH81N binds peptides to form complexes similar to peptide–DR1WT.
Figure 2: Kinetics of dissociation of peptides in the absence of DM are faster for mutant DR1βH81N and resemble DM-mediated peptide dissociation.
Figure 3: Pocket 1 of HA(306–318)–DR1.
Figure 4: DM has a minimal effect on the dissociation of peptide from DR1βH81N.
Figure 5: The βH81N substitution does not alter the interaction of DM with DR.
Figure 6: Reintroduction of an appropriate histidine residue partially restores DM-mediated dissociation.

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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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Author notes

  1. Kedar Narayan and Chih-Ling Chou: These authors contributed equally to this work.

Authors and Affiliations

  1. Graduate Program in Immunology, Johns Hopkins University, Baltimore, 21205, Maryland, USA

    Kedar Narayan, AeRyon Kim, Isamu Z Hartman & Scheherazade Sadegh-Nasseri

  2. Graduate Program in Molecular and Computational Biophysics, Johns Hopkins University, Baltimore, 21205, Maryland, USA

    Chih-Ling Chou & Scheherazade Sadegh-Nasseri

  3. Department of Pathology, Johns Hopkins University, Baltimore, 21205, Maryland, USA

    Sarat Dalai, Stanislav Khoruzhenko & Scheherazade Sadegh-Nasseri

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Correspondence to Scheherazade Sadegh-Nasseri.

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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)

Supplementary Note (PDF 51 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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