title: Engineering of metal-ion sites as distance constraints in structural and functional analysis of 7TM receptors.
abstract: G-protein-coupled receptors with their seven transmembrane (7TM) segments constitute the largest superfamily of proteins known. Unfortunately, still only relatively low resolution structures derived from electron cryo-microscopy analysis of 2D crystals are available for these proteins. We have used artificially designed Zn(II) metal-ion binding sites to probe 7TM receptors structurally and functionally and to define some basic distance constraints for molecular modeling. In this way, the relative helical rotation and vertical translocation of transmembrane helices TM-II, TM-III, TM-V, and TM-VI of the tachykinin NK-1 receptor have been restricted. Collectively, these zinc sites constitute a basic network of distance constraints that limit the degrees of freedom of the interhelical contact faces in molecular models of 7TM receptors. The construction of artificially designed metal-ion sites is discussed also in the context of probes for conformational changes occurring during receptor activation.
authors.affiliation: Laboratory for Molecular Pharmacology, Rigshospitalet, University of Copenhagen, Denmark. elling@molpharm.dk
authors.forename: C E
authors.initials: CE
authors.lastname: Elling
authors.forename: K
authors.initials: K
authors.lastname: Thirstrup
authors.forename: S M
authors.initials: SM
authors.lastname: Nielsen
authors.forename: S A
authors.initials: SA
authors.lastname: Hjorth
authors.forename: T W
authors.initials: TW
authors.lastname: Schwartz
mesh_terms: D001665:Binding Sites; D002413:Cations, Divalent; D019204:GTP-Binding Proteins / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000737:chemistry* / Q000378:metabolism; D013329:Structure-Activity Relationship; D015032:Zinc / Q000378:metabolism
chemical_list: D002413:Cations, Divalent; D011956:Receptors, Cell Surface; D019204:GTP-Binding Proteins; D015032:Zinc