title: Detecting molecular interactions that stabilize native bovine rhodopsin.
abstract: Using single-molecule force spectroscopy we probed molecular interactions within native bovine rhodopsin and discovered structural segments of well-defined mechanical stability. Highly conserved residues among G protein-coupled receptors were located at the interior of individual structural segments, suggesting a dual role for these segments in rhodopsin. Firstly, structural segments stabilize secondary structure elements of the native protein, and secondly, they position and hold the highly conserved residues at functionally important environments. Two main classes of force curves were observed. One class corresponded to the unfolding of rhodopsin with the highly conserved Cys110-Cys187 disulfide bond remaining intact and the other class corresponded to the unfolding of the entire rhodopsin polypeptide chain. In the absence of the Cys110-Cys187 bond, the nature of certain molecular interactions within folded rhodopsin was altered. These changes highlight the structural importance of this disulfide bond and may form the basis of dysfunctions associated with its absence.
authors.affiliation: Center for Biotechnology, University of Technology, 01307 Dresden, Germany.
authors.forename: K
authors.initials: K
authors.lastname: Tanuj Sapra
authors.forename: Paul S-H
authors.initials: PS
authors.lastname: Park
authors.forename: Slawomir
authors.initials: S
authors.lastname: Filipek
authors.forename: Andreas
authors.initials: A
authors.lastname: Engel
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Müller
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Palczewski
mesh_terms: D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000648:ultrastructure; D003545:Cysteine / Q000378:metabolism; D003624:Darkness; D021621:Imaging, Three-Dimensional; D008958:Models, Molecular; D008961:Models, Structural; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D012243:Rhodopsin / Q000737:chemistry* / Q000378:metabolism*; D012374:Rod Cell Outer Segment / Q000648:ultrastructure; D013816:Thermodynamics
chemical_list: D012243:Rhodopsin; D003545:Cysteine