title: Preliminary X-ray crystallographic analysis of 2-methylcitrate synthase from Salmonella typhimurium.
abstract: 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.
authors.affiliation: Molecular Biophysics Unit, Indian Institute of Science, Bangalore, Karnataka 560012, India.
authors.forename: Sagar
authors.initials: S
authors.lastname: Chittori
authors.forename: D K
authors.initials: DK
authors.lastname: Simanshu
authors.forename: H S
authors.initials: HS
authors.lastname: Savithri
authors.forename: M R N
authors.initials: MR
authors.lastname: Murthy
mesh_terms: D018360:Crystallography, X-Ray; D007652:Oxo-Acid-Lyases / Q000737:chemistry* / Q000302:isolation & purification; D012486:Salmonella typhimurium / Q000201:enzymology*
chemical_list: C112399:2-methylcitrate synthase; D007652:Oxo-Acid-Lyases
references.citation: Biochemistry. 2001 Nov 6;40(44):13177-87
references.pmid: 11683626
references.citation: Eur J Biochem. 2002 Dec;269(24):6250-60
references.pmid: 12473121
references.citation: J Mol Biol. 1982 Jun 15;158(1):111-52
references.pmid: 7120407
references.citation: J Mol Biol. 1984 Mar 25;174(1):205-19
references.pmid: 6716477
references.citation: Methods Enzymol. 1997;276:307-26
references.pmid: 27754618
references.citation: Methods Enzymol. 1997;276:594-611
references.pmid: 9048382
references.citation: J Biol Chem. 2001 Jun 1;276(22):19094-101
references.pmid: 11376009
references.citation: Adv Microb Physiol. 1991;32:87-108
references.pmid: 1882730
references.citation: Biochem J. 1964 Aug;92(2):338-45
references.pmid: 4284461
references.citation: J Mol Biol. 1968 Apr 28;33(2):491-7
references.pmid: 5700707
references.citation: Biochemistry. 1997 Aug 19;36(33):9983-94
references.pmid: 9254593
references.citation: Microbiology (Reading). 2007 Dec;153(Pt 12):3973-3982
references.pmid: 18048912
references.citation: Curr Top Cell Regul. 1992;33:209-29
references.pmid: 1499334
references.citation: J Mol Biol. 2007 May 18;368(5):1332-44
references.pmid: 17395198
references.citation: J Hyg (Lond). 1979 Apr;82(2):263-83
references.pmid: 34658
references.citation: J Infect Dis. 1962 Sep-Oct;111:117-27
references.pmid: 13968487
references.citation: Biochemistry. 2006 Nov 14;45(45):13487-99
references.pmid: 17087502
references.citation: J Mol Biol. 2003 May 2;328(3):609-21
references.pmid: 12706720
references.citation: J Biol Chem. 1977 Apr 25;252(8):2702-9
references.pmid: 856801
references.citation: Biochem Biophys Res Commun. 2003 Nov 7;311(1):193-201
references.pmid: 14575713
references.citation: J Bacteriol. 1997 Feb;179(3):928-40
references.pmid: 9006051
references.citation: Pediatr Res. 1975 Dec;9(12):905-8
references.pmid: 127973
references.citation: Biochemistry. 2001 Apr 17;40(15):4703-13
references.pmid: 11294638
references.citation: Mol Microbiol. 2006 Jun;60(5):1109-22
references.pmid: 16689789
references.citation: Biochem J. 1964 Mar;90(3):483-9
references.pmid: 5834509
references.citation: Gut. 1987 Oct;28(10):1221-7
references.pmid: 3678950
references.citation: Arch Microbiol. 1997 Nov;168(5):428-36
references.pmid: 9325432
references.citation: FEMS Microbiol Lett. 1996 Apr 1;137(2-3):233-9
references.pmid: 8998991
references.citation: Structure. 1998 Mar 15;6(3):351-61
references.pmid: 9551556