title: Calreticulin is a thermostable protein with distinct structural responses to different divalent cation environments.
abstract: Calreticulin is a soluble calcium-binding chaperone of the endoplasmic reticulum (ER) that is also detected on the cell surface and in the cytosol. Calreticulin contains a single high affinity calcium-binding site within a globular domain and multiple low affinity sites within a C-terminal acidic region. We show that the secondary structure of calreticulin is remarkably thermostable at a given calcium concentration. Rather than corresponding to complete unfolding events, heat-induced structural transitions observed for calreticulin relate to tertiary structural changes that expose hydrophobic residues and reduce protein rigidity. The thermostability and the overall secondary structure content of calreticulin are impacted by the divalent cation environment, with the ER range of calcium concentrations enhancing stability, and calcium-depleting or high calcium environments reducing stability. Furthermore, magnesium competes with calcium for binding to calreticulin and reduces thermostability. The acidic domain of calreticulin is an important mediator of calcium-dependent changes in secondary structure content and thermostability. Together, these studies indicate interactions between the globular and acidic domains of calreticulin that are impacted by divalent cations. These interactions influence the structure and stability of calreticulin, and are likely to determine the multiple functional activities of calreticulin in different subcellular environments.
authors.affiliation: Department of Microbiology and Immunology, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Sanjeeva J
authors.initials: SJ
authors.lastname: Wijeyesakere
authors.forename: Ari A
authors.initials: AA
authors.lastname: Gafni
authors.forename: Malini
authors.initials: M
authors.lastname: Raghavan
mesh_terms: D000818:Animals; D064032:Calbindin 2; D002118:Calcium / Q000737:chemistry*; D002413:Cations, Divalent / Q000737:chemistry*; D006358:Hot Temperature; D008274:Magnesium / Q000737:chemistry*; D051379:Mice; D017510:Protein Folding; D055550:Protein Stability; D017434:Protein Structure, Tertiary; D064030:S100 Calcium Binding Protein G / Q000737:chemistry*; D013329:Structure-Activity Relationship
chemical_list: D064032:Calbindin 2; D002413:Cations, Divalent; D064030:S100 Calcium Binding Protein G; D008274:Magnesium; D002118:Calcium