title: Characterization of the human sigma-1 receptor chaperone domain structure and binding immunoglobulin protein (BiP) interactions.
abstract: The sigma-1 receptor (S1R) is a ligand-regulated membrane protein chaperone involved in the ER stress response. S1R activity is implicated in diseases of the central nervous system including amnesia, schizophrenia, depression, Alzheimer disease, and addiction. S1R has been shown previously to regulate the Hsp70 binding immunoglobulin protein (BiP) and the inositol triphosphate receptor calcium channel through a C-terminal domain. We have developed methods for bacterial expression and reconstitution of the chaperone domain of human S1R into detergent micelles that enable its study by solution NMR spectroscopy. The chaperone domain is found to contain a helix at the N terminus followed by a largely dynamic region and a structured, helical C-terminal region that encompasses a membrane associated domain containing four helices. The helical region at residues ∼198-206 is strongly amphipathic and proposed to anchor the chaperone domain to micelles and membranes. Three of the helices in the C-terminal region closely correspond to previously identified cholesterol and drug recognition sites. In addition, it is shown that the chaperone domain interacts with full-length BiP or the isolated nucleotide binding domain of BiP, but not the substrate binding domain, suggesting that the nucleotide binding domain is sufficient for S1R interactions.
authors.affiliation: From the Department of Biochemistry, University of Oxford, Oxford OX1 3QU, United Kingdom.
authors.forename: Jose Luis
authors.initials: JL
authors.lastname: Ortega-Roldan
authors.affiliation: From the Department of Biochemistry, University of Oxford, Oxford OX1 3QU, United Kingdom.
authors.forename: Felipe
authors.initials: F
authors.lastname: Ossa
authors.affiliation: From the Department of Biochemistry, University of Oxford, Oxford OX1 3QU, United Kingdom. Electronic address: jason.schnell@bioch.ox.ac.uk.
authors.forename: Jason R
authors.initials: JR
authors.lastname: Schnell
mesh_terms: D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D000091342:Endoplasmic Reticulum Chaperone BiP; D006360:Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008823:Micelles; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011522:Protons; D017480:Receptors, sigma / Q000737:chemistry* / Q000378:metabolism*; D014867:Water / Q000737:chemistry; D000097605:Sigma-1 Receptor
chemical_list: D000091342:Endoplasmic Reticulum Chaperone BiP; D006360:Heat-Shock Proteins; D008823:Micelles; D011522:Protons; D017480:Receptors, sigma; D014867:Water; D000097605:Sigma-1 Receptor
keywords: BiP; Endoplasmic Reticulum Stress; Membrane Proteins; NMR; Sigma-1 Receptor; Signaling; Structural Biology