title: Purification, crystallization and preliminary X-ray analysis of the aspartate aminotransferase of Plasmodium falciparum.
abstract: Aspartate aminotransferases (EC 2.6.1.1) catalyse the conversion of aspartate and alpha-ketoglutarate to oxaloacetate and glutamate in a reversible manner. Thus, the aspartate aminotransferase of Plasmodium falciparum (PfAspAT) plays a central role in the transamination of amino acids. Recent findings suggest that PfAspAT may also play a pivotal role in energy metabolism and the de novo biosynthesis of pyrimidines. While therapeutics based upon the inhibition of other proteins in these pathways are already used in the treatment of malaria, the advent of multidrug-resistant strains has limited their efficacy. The presence of PfAspAT in these pathways may offer additional opportunities for the development of novel therapeutics. In order to gain a deeper understanding of the function and role of PfAspAT, it has been expressed and purified to homogeneity. The successful crystallization of PfAspAT, the collection of a 2.8 A diffraction data set and initial attempts to solve the structure using molecular replacement are reported.
authors.affiliation: European Molecular Biology Laboratory Hamburg Outstation, Notkestrasse 85, D-22603 Hamburg, Germany.
authors.forename: Rishabh
authors.initials: R
authors.lastname: Jain
authors.forename: Rositsa
authors.initials: R
authors.lastname: Jordanova
authors.forename: Ingrid B
authors.initials: IB
authors.lastname: Müller
authors.forename: Carsten
authors.initials: C
authors.lastname: Wrenger
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Groves
mesh_terms: D001219:Aspartate Aminotransferases / Q000737:chemistry* / Q000302:isolation & purification; D003460:Crystallization; D018360:Crystallography, X-Ray; D010963:Plasmodium falciparum / Q000201:enzymology*
chemical_list: D001219:Aspartate Aminotransferases
references.citation: Lancet Infect Dis. 2002 Sep;2(9):564-73
references.pmid: 12206972
references.citation: Biochemistry. 2007 Sep 18;46(37):10517-27
references.pmid: 17713924
references.citation: J Bacteriol. 2001 Aug;183(15):4421-34
references.pmid: 11443076
references.citation: J Biochem. 1996 Jan;119(1):135-44
references.pmid: 8907187
references.citation: N Engl J Med. 2009 Jul 30;361(5):455-67
references.pmid: 19641202
references.citation: J Bacteriol. 1999 Mar;181(6):1739-47
references.pmid: 10074065
references.citation: Lancet Infect Dis. 2002 Apr;2(4):209-18
references.pmid: 11937421
references.citation: Antimicrob Agents Chemother. 2000 Sep;44(9):2540-2
references.pmid: 10952611
references.citation: Annu Rev Pharmacol Toxicol. 1995;35:55-91
references.pmid: 7598507
references.citation: Antimicrob Agents Chemother. 1995 Nov;39(11):2511-5
references.pmid: 8585735
references.citation: Eur J Biochem. 1993 Jun 1;214(2):549-61
references.pmid: 8513804
references.citation: Acta Crystallogr D Biol Crystallogr. 2007 Apr;63(Pt 4):526-35
references.pmid: 17372358
references.citation: Acta Crystallogr D Biol Crystallogr. 2006 Jan;62(Pt 1):72-82
references.pmid: 16369096
references.citation: Nature. 1970 Aug 15;227(5259):680-5
references.pmid: 5432063
references.citation: J Biol Chem. 2005 Mar 11;280(10):9547-54
references.pmid: 15576366
references.citation: Acta Crystallogr D Biol Crystallogr. 2006 Dec;62(Pt 12):1446-52
references.pmid: 17139079
references.citation: FEMS Microbiol Lett. 1998 Aug 15;165(2):305-12
references.pmid: 9742703
references.citation: N Engl J Med. 2009 Jul 30;361(5):540-1
references.pmid: 19641219
references.citation: J Indian Med Assoc. 2008 Oct;106(10):678-81, 683
references.pmid: 19552104
references.citation: Eur J Biochem. 1990 Apr 20;189(1):1-23
references.pmid: 2185018
references.citation: Pharmacol Ther. 2001 Feb;89(2):207-19
references.pmid: 11316521
references.citation: Nucleic Acids Res. 2000 Jan 1;28(1):235-42
references.pmid: 10592235
references.citation: J Antimicrob Chemother. 2003 Aug;52(2):290-3
references.pmid: 12837742
references.citation: Acta Crystallogr D Biol Crystallogr. 2008 Jan;64(Pt 1):125-32
references.pmid: 18094476
references.citation: Protein Sci. 2003 Sep;12(9):1865-71
references.pmid: 12930986
references.citation: Antimicrob Agents Chemother. 2005 Jul;49(7):2857-64
references.pmid: 15980361
references.citation: Acta Crystallogr D Biol Crystallogr. 2006 Jan;62(Pt 1):48-57
references.pmid: 16369093
references.citation: Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
references.pmid: 15572765
references.citation: Methods Mol Biol. 2008;426:299-318
references.pmid: 18542872
references.citation: Acta Crystallogr D Biol Crystallogr. 2006 Jan;62(Pt 1):58-64
references.pmid: 16369094
references.citation: Am J Trop Med Hyg. 2009 Aug;81(2):338-42
references.pmid: 19635895
references.citation: J Mol Biol. 1968 Apr 28;33(2):491-7
references.pmid: 5700707
references.citation: Mol Biochem Parasitol. 2008 Jul;160(1):1-7
references.pmid: 18455248
references.citation: J Biol Chem. 1998 Feb 27;273(9):4982-9
references.pmid: 9478945
references.citation: Acta Crystallogr D Biol Crystallogr. 2003 Jul;59(Pt 7):1145-53
references.pmid: 12832757