Open AccessOverproduction, purification, crystallization and preliminary X‐ray diffraction analysis of Trypanosoma brucei gambiense glycerol kinase
Author(s) -
Balogun Emmanuel Oluwadare,
Inaoka Daniel Ken,
Kido Yasutoshi,
Shiba Tomoo,
Nara Takeshi,
Aoki Takashi,
Honma Teruki,
Tanaka Akiko,
Inoue Masayuki,
Matsuoka Shigeru,
Michels Paul A. M.,
Harada Shigeharu,
Kita Kiyoshi
Publication year - 2010
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309110000369
Subject(s) - trypanosoma brucei , glycerol kinase , orthorhombic crystal system , glycerol , crystallization , enzyme , chemistry , kinase , solvent , crystallography , biochemistry , crystal structure , organic chemistry , gene
In the bloodstream forms of human trypanosomes, glycerol kinase (GK; EC 2.7.1.30) is one of the nine glycosomally compartmentalized enzymes that are essential for energy metabolism. In this study, a recombinant Trypanosoma brucei gambiense GK (rTbgGK) with an N‐terminal cleavable His 6 tag was overexpressed, purified to homogeneity and crystallized by the sitting‐drop vapour‐diffusion method using PEG 400 as a precipitant. A complete X‐ray diffraction data set to 2.75 Å resolution indicated that the crystals belonged to the orthorhombic space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 63.84, b = 121.50, c = 154.59 Å. The presence of two rTbgGK molecules in the asymmetric unit gives a Matthews coefficient ( V M ) of 2.5 Å 3 Da −1 , corresponding to 50% solvent content.