Open AccessCrystallization and preliminary X‐ray crystallographic analysis of the receptor‐uncoupled mutant of Gα i1
Author(s) -
Morikawa Tomohito,
Muroya Ayumu,
Nakajima Yoshitaka,
Tanaka Takeshi,
Hirai Keiko,
Sugio Shigetoshi,
Kohno Toshiyuki,
Wakamatsu Kaori
Publication year - 2007
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309107003363
Subject(s) - gtp' , crystallization , crystallography , mutant , g protein coupled receptor , g protein , chemistry , receptor , muscarinic acetylcholine receptor , x ray crystallography , diffraction , biophysics , biology , biochemistry , physics , gene , optics , organic chemistry , enzyme
In order to understand the molecular mechanisms by which G‐protein‐coupled receptors (GPCRs) activate G proteins, the K349P mutant of Gα 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 Gα 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 GTPγS) and the crystals diffracted to resolutions of 2.2 Å (WT·GDP), 2.8 Å (WT·GTPγS), 2.6 Å (K349P·GDP) and 3.2 Å (K349P·GTPγS).