title: Cloning, expression, purification and preliminary X-ray crystallographic studies of 2-methylisocitrate lyase from Salmonella typhimurium.
abstract: In Salmonella typhimurium, propionate is oxidized to pyruvate via the 2-methylcitric acid cycle. The last step of this cycle, the cleavage of 2-methylisocitrate to succinate and pyruvate, is catalysed by 2-methylisocitrate lyase (EC 4.1.3.30). Methylisocitrate lyase (molecular weight 32 kDa) with a C-terminal polyhistidine affinity tag has been cloned and overexpressed in Escherichia coli and purified and crystallized under different conditions using the hanging-drop vapour-diffusion technique. Crystals belong to the orthogonal space group P2(1)2(1)2(1), with unit-cell parameters a = 63.600, b = 100.670, c = 204.745 A. A complete data set to 2.5 A resolution has been collected using an image-plate detector system mounted on a rotating-anode X-ray generator.
authors.affiliation: Molecular Biophysics Unit, Indian Institute of Science, Bangalore, India.
authors.forename: Dhirendra K
authors.initials: DK
authors.lastname: Simanshu
authors.forename: P S
authors.initials: PS
authors.lastname: Satheshkumar
authors.forename: S
authors.initials: S
authors.lastname: Parthasarathy
authors.forename: H S
authors.initials: HS
authors.lastname: Savithri
authors.forename: M R N
authors.initials: MR
authors.lastname: Murthy
mesh_terms: D001483:Base Sequence; D019755:Carbon-Carbon Lyases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D017931:DNA Primers; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D012486:Salmonella typhimurium / Q000201:enzymology*
chemical_list: D017931:DNA Primers; D011994:Recombinant Proteins; D019755:Carbon-Carbon Lyases; C417277:methylisocitrate lyase