Preliminary X-ray crystallographic analysis of 2-methylcitrate synthase from Salmonella typhimurium.
Analysis of the genomic sequences of Escherichia coli and Salmonella typhimurium has revealed the presence of several homologues of the well studied citrate synthase (CS). One of these homologues has been shown to code for 2-methylcitrate synthase (2-MCS) activity. 2-MCS catalyzes one of the steps in the 2-methylcitric acid cycle found in these organisms for the degradation of propionate to pyruvate and succinate. In the present work, the gene coding for 2-MCS from S. typhimurium (StPrpC) was cloned in pRSET-C vector and overexpressed in E. coli. The protein was purified to homogeneity using Ni-NTA affinity chromatography. The purified protein was crystallized using the microbatch-under-oil method. The StPrpC crystals diffracted X-rays to 2.4 A resolution and belonged to the triclinic space group P1, with unit-cell parameters a = 92.068, b = 118.159, c = 120.659 A, alpha = 60.84, beta = 67.77, gamma = 81.92 degrees . Computation of rotation functions using the X-ray diffraction data shows that the protein is likely to be a decamer of identical subunits, unlike CSs, which are dimers or hexamers.
Chittori
Sagar
S
Molecular Biophysics Unit, Indian Institute of Science, Bangalore, Karnataka 560012, India.
Simanshu
D K
DK
Savithri
H S
HS
Murthy
M R N
MR
D018360:Crystallography, X-Ray; D007652:Oxo-Acid-Lyases / Q000737:chemistry* / Q000302:isolation & purification; D012486:Salmonella typhimurium / Q000201:enzymology*
C112399:2-methylcitrate synthase; D007652:Oxo-Acid-Lyases
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