title: Cloning, expression, purification, crystallization and initial crystallographic analysis of the lysine-biosynthesis LysX protein from Thermus thermophilus HB8.
abstract: The gene encoding LysX, an essential component of the lysine-biosynthesis pathway in Thermus thermophilus (molecular weight approximately equal 31,000 Da), was cloned and expressed and the purified protein was crystallized by the hanging-drop vapour-diffusion technique in two different space groups, C2 (unit-cell parameters a = 124.7, b = 51.4, c = 103.6 A, beta = 122.8 degrees ) and R3 (a = b = 122.6, c = 97.6 A). Crystals improved by macroseeding diffracted to beyond 2.3 and 3 A resolution for the C2 and R3 crystal forms, respectively. Complete diffraction data sets were collected for the C2 and R3 crystal forms at 2.5 and 3.1 A resolution, respectively. Crystals of selenomethionine-containing LysX protein were obtained by cross-microseeding, using the native microcrystals as a seed. Structure determination is now in progress.
authors.affiliation: Genomic Sciences Center, RIKEN Yokohama Institute, 1-7-22 Suehiro-cho, Tsurumi, Yokohama 230-0045, Japan.
authors.forename: Marina N
authors.initials: MN
authors.lastname: Vassylyeva
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Sakai
authors.forename: Takanori
authors.initials: T
authors.lastname: Matsuura
authors.forename: Shun-ichi
authors.initials: S
authors.lastname: Sekine
authors.forename: Makoto
authors.initials: M
authors.lastname: Nishiyama
authors.forename: Takaho
authors.initials: T
authors.lastname: Terada
authors.forename: Mikako
authors.initials: M
authors.lastname: Shirouzu
authors.forename: Seiki
authors.initials: S
authors.lastname: Kuramitsu
authors.forename: Dmitry G
authors.initials: DG
authors.lastname: Vassylyev
authors.forename: Shigeyuki
authors.initials: S
authors.lastname: Yokoyama
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry*; D003001:Cloning, Molecular / Q000379:methods; D003460:Crystallization / Q000379:methods*; D018360:Crystallography, X-Ray; D008239:Lysine / Q000096:biosynthesis / Q000737:chemistry*; D016963:Thermus thermophilus / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; C477978:LysX protein, T thermophilus; D008239:Lysine