title: Overproduction, purification, crystallization and preliminary X-ray diffraction analysis of Trypanosoma brucei gambiense glycerol kinase.
abstract: 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 A resolution indicated that the crystals belonged to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 63.84, b = 121.50, c = 154.59 A. The presence of two rTbgGK molecules in the asymmetric unit gives a Matthews coefficient (V(M)) of 2.5 A(3) Da(-1), corresponding to 50% solvent content.
authors.affiliation: Department of Biomedical Chemistry, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Emmanuel Oluwadare
authors.initials: EO
authors.lastname: Balogun
authors.forename: Daniel Ken
authors.initials: DK
authors.lastname: Inaoka
authors.forename: Yasutoshi
authors.initials: Y
authors.lastname: Kido
authors.forename: Tomoo
authors.initials: T
authors.lastname: Shiba
authors.forename: Takeshi
authors.initials: T
authors.lastname: Nara
authors.forename: Takashi
authors.initials: T
authors.lastname: Aoki
authors.forename: Teruki
authors.initials: T
authors.lastname: Honma
authors.forename: Akiko
authors.initials: A
authors.lastname: Tanaka
authors.forename: Masayuki
authors.initials: M
authors.lastname: Inoue
authors.forename: Shigeru
authors.initials: S
authors.lastname: Matsuoka
authors.forename: Paul A M
authors.initials: PA
authors.lastname: Michels
authors.forename: Shigeharu
authors.initials: S
authors.lastname: Harada
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Kita
mesh_terms: D000818:Animals; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003460:Crystallization; D018360:Crystallography, X-Ray; D005991:Glycerol Kinase / Q000737:chemistry* / Q000302:isolation & purification; D014347:Trypanosoma brucei gambiense / Q000201:enzymology*
chemical_list: D005991:Glycerol Kinase