title: Interaction of HLA-DR with an acidic face of HLA-DM disrupts sequence-dependent interactions with peptides.
abstract: HLA-DM (DM) edits major histocompatibility complex class II (MHCII)-bound peptides in endocytic compartments and stabilizes empty MHCII molecules. Crystal structures of DM have revealed similarity to MHCII but not how DM and MHCII interact. We used mutagenesis to map a MHCII-interacting surface on DM. Mutations on this surface impair DM action on HLA-DR and -DP in cells and DM-dependent peptide loading in vitro. The orientation of DM and MHCII predicted by these studies guided design of soluble DM and DR molecules fused to leucine zippers via their beta chains, resulting in stable DM/DR complexes. Peptide release from the complexes was fast and only weakly sequence dependent, arguing that DM diminishes the selectivity of the MHCII groove. Analysis of soluble DM action on soluble DR/peptide complexes corroborates this conclusion.
authors.affiliation: Department of Pediatrics, Stanford University, Stanford, California 94305, USA.
authors.forename: Achal
authors.initials: A
authors.lastname: Pashine
authors.forename: Robert
authors.initials: R
authors.lastname: Busch
authors.forename: Michael P
authors.initials: MP
authors.lastname: Belmares
authors.forename: Jason N
authors.initials: JN
authors.lastname: Munning
authors.forename: Robert C
authors.initials: RC
authors.lastname: Doebele
authors.forename: Megan
authors.initials: M
authors.lastname: Buckingham
authors.forename: Gary P
authors.initials: GP
authors.lastname: Nolan
authors.forename: Elizabeth D
authors.initials: ED
authors.lastname: Mellins
mesh_terms: D000944:Antigens, Differentiation, B-Lymphocyte / Q000378:metabolism; D001483:Base Sequence; D002460:Cell Line; D018076:DNA, Complementary / Q000235:genetics; D004355:Drug Stability; D006681:HLA-D Antigens / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006684:HLA-DR Antigens / Q000737:chemistry / Q000378:metabolism*; D000949:Histocompatibility Antigens Class II / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D046911:Macromolecular Substances; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D012995:Solubility
chemical_list: D000944:Antigens, Differentiation, B-Lymphocyte; D018076:DNA, Complementary; D006681:HLA-D Antigens; C084245:HLA-DM antigens; D006684:HLA-DR Antigens; D000949:Histocompatibility Antigens Class II; D046911:Macromolecular Substances; C045084:invariant chain