Crystallization and preliminary X-ray crystallographic analysis of the receptor-uncoupled mutant of Galphai1.
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).
Morikawa
Tomohito
T
Mitsubishi Kagaku Institute of Life Sciences (MITILS), 11 Minamiooya, Machida, Tokyo 194-8511, Japan.
Muroya
Ayumu
A
Nakajima
Yoshitaka
Y
Tanaka
Takeshi
T
Hirai
Keiko
K
Sugio
Shigetoshi
S
Wakamatsu
Kaori
K
Kohno
Toshiyuki
T
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
D044385:GTP-Binding Protein alpha Subunits; D043562:Receptors, G-Protein-Coupled