Empty class II major histocompatibility complex created by peptide photolysis establishes the role of DM in peptide association

Gijsbert M. Grotenbreg, Melissa J. Nicholson, Kevin D. Fowler, Kathrin Wilbuer, Leah Octavio, Maxine Yang, Arup K. Chakraborty, Hidde L. Ploegh, Kai W. Wucherpfennig

Research output: Contribution to journalArticlepeer-review

45 Scopus citations

Abstract

DM catalyzes the exchange of peptides bound to Class II major histocompatibility complex (MHC) molecules. Because the dissociation and association components of the overall reaction are difficult to separate, a detailed mechanism of DM catalysis has long resisted elucidation. UV irradiation of DR molecules loaded with a photocleavable peptide (caged Class IIMHC molecules) enabled synchronous and verifiable evacuation of the peptide-binding groove and tracking of early binding events in real time by fluorescence polarization. Empty DR molecules generated by photocleavage rapidly bound peptide but quickly resolved into species with substantially slower binding kinetics. DM formed a complex with empty DR molecules that bound peptide with even faster kinetics than empty DR molecules just having lost their peptide cargo. Mathematical models demonstrate that the peptide association rate of DR molecules is substantially higher in the presence of DM. We therefore unequivocally establish that DM contributes directly to peptide association through formation of a peptide-loading complex between DM and empty Class II MHC. This complex rapidly acquires a peptide analogous to the MHC class I peptide-loading complex.

Original languageEnglish
Pages (from-to)21425-21436
Number of pages12
JournalJournal of Biological Chemistry
Volume282
Issue number29
DOIs
StatePublished - 20 Jul 2007
Externally publishedYes

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