title: Regulation of calreticulin-major histocompatibility complex (MHC) class I interactions by ATP.
abstract: The MHC class I peptide loading complex (PLC) facilitates the assembly of MHC class I molecules with peptides, but factors that regulate the stability and dynamics of the assembly complex are largely uncharacterized. Based on initial findings that ATP, in addition to MHC class I-specific peptide, is able to induce MHC class I dissociation from the PLC, we investigated the interaction of ATP with the chaperone calreticulin, an endoplasmic reticulum (ER) luminal, calcium-binding component of the PLC that is known to bind ATP. We combined computational and experimental measurements to identify residues within the globular domain of calreticulin, in proximity to the high-affinity calcium-binding site, that are important for high-affinity ATP binding and for ATPase activity. High-affinity calcium binding by calreticulin is required for optimal nucleotide binding, but both ATP and ADP destabilize enthalpy-driven high-affinity calcium binding to calreticulin. ATP also selectively destabilizes the interaction of calreticulin with cellular substrates, including MHC class I molecules. Calreticulin mutants that affect ATP or high-affinity calcium binding display prolonged associations with monoglucosylated forms of cellular MHC class I, delaying MHC class I dissociation from the PLC and their transit through the secretory pathway. These studies reveal central roles for ATP and calcium binding as regulators of calreticulin-substrate interactions and as key determinants of PLC dynamics.
authors.affiliation: Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, MI 48109;
authors.forename: Sanjeeva Joseph
authors.initials: SJ
authors.lastname: Wijeyesakere
authors.affiliation: Department of Chemistry, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Jessica K
authors.initials: JK
authors.lastname: Gagnon
authors.affiliation: Department of Chemistry, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Karunesh
authors.initials: K
authors.lastname: Arora
authors.affiliation: Department of Chemistry, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Charles L
authors.initials: CL
authors.lastname: Brooks
authors.affiliation: Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, MI 48109; malinir@umich.edu.
authors.forename: Malini
authors.initials: M
authors.lastname: Raghavan
mesh_terms: D000255:Adenosine Triphosphate / Q000235:genetics / Q000276:immunology*; D000818:Animals; D064032:Calbindin 2 / Q000235:genetics / Q000276:immunology*; D002118:Calcium / Q000276:immunology; D004721:Endoplasmic Reticulum / Q000235:genetics / Q000276:immunology*; D015395:Histocompatibility Antigens Class I / Q000235:genetics / Q000276:immunology*; D051379:Mice; D018345:Mice, Knockout; D011485:Protein Binding / Q000235:genetics / Q000276:immunology; D017434:Protein Structure, Tertiary
chemical_list: C576511:Calb2 protein, mouse; D064032:Calbindin 2; D015395:Histocompatibility Antigens Class I; D000255:Adenosine Triphosphate; D002118:Calcium
keywords: ATP; ATPase; MHC class; calreticulin; monoglucosylated glycans