Empty class II major histocompatibility complex created by peptide photolysis establishes the role of DM in peptide association.
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 II MHC 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.
Grotenbreg
Gijsbert M
GM
Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02139, USA.
Nicholson
Melissa J
MJ
Fowler
Kevin D
KD
Wilbuer
Kathrin
K
Octavio
Leah
L
Yang
Maxine
M
Chakraborty
Arup K
AK
Ploegh
Hidde L
HL
Wucherpfennig
Kai W
KW
D006681:HLA-D Antigens / Q000737:chemistry* / Q000502:physiology; D000949:Histocompatibility Antigens Class II / Q000235:genetics*; D006801:Humans; D007700:Kinetics; D008027:Light; D008285:Major Histocompatibility Complex; D008954:Models, Biological; D008956:Models, Chemical; D008962:Models, Theoretical; D010455:Peptides / Q000737:chemistry*; D010777:Photochemistry / Q000379:methods; D010782:Photolysis; D011485:Protein Binding; D013997:Time Factors; D014466:Ultraviolet Rays
D006681:HLA-D Antigens; C084245:HLA-DM antigens; D000949:Histocompatibility Antigens Class II; D010455:Peptides