title: Crystallization and preliminary X-ray crystallographic analysis of the receptor-uncoupled mutant of Galphai1.
abstract: In order to understand the molecular mechanisms by which G-protein-coupled receptors (GPCRs) activate G proteins, the K349P mutant of Galpha(i1) (K349P), which is unable to couple to the muscarinic acetylcholine receptor, was prepared and its crystals were grown along with those of wild-type Galpha(i1) protein (WT). The two proteins were crystallized under almost identical conditions, thus enabling a detailed structural comparison. The crystallization conditions performed well irrespective of the identity of the bound nucleotide (GDP or GTPgammaS) and the crystals diffracted to resolutions of 2.2 A (WT.GDP), 2.8 A (WT.GTPgammaS), 2.6 A (K349P.GDP) and 3.2 A (K349P.GTPgammaS).
authors.affiliation: Mitsubishi Kagaku Institute of Life Sciences (MITILS), 11 Minamiooya, Machida, Tokyo 194-8511, Japan.
authors.forename: Tomohito
authors.initials: T
authors.lastname: Morikawa
authors.forename: Ayumu
authors.initials: A
authors.lastname: Muroya
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Nakajima
authors.forename: Takeshi
authors.initials: T
authors.lastname: Tanaka
authors.forename: Keiko
authors.initials: K
authors.lastname: Hirai
authors.forename: Shigetoshi
authors.initials: S
authors.lastname: Sugio
authors.forename: Kaori
authors.initials: K
authors.lastname: Wakamatsu
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Kohno
mesh_terms: D000818:Animals; D003460:Crystallization; D018360:Crystallography, X-Ray; D044385:GTP-Binding Protein alpha Subunits / Q000737:chemistry* / Q000235:genetics; D009154:Mutation; D051381:Rats; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry* / Q000235:genetics
chemical_list: D044385:GTP-Binding Protein alpha Subunits; D043562:Receptors, G-Protein-Coupled