title: Backbone and ILV methyl resonance assignments of E. coli thymidylate synthase bound to cofactor and a nucleotide analogue.
abstract: Thymidylate synthase (TSase) is a 62 kDa homodimeric enzyme required for de novo synthesis of thymidine monophosphate in most organisms. This makes the enzyme an excellent target for anticancer and microbial antibiotic drugs. In addition, TSase has been shown to exhibit negative cooperativity and half-the-sites reactivity. For these collective reasons, TSase is widely studied, and much is known about its kinetics and structure as it progresses through a multi-step catalytic cycle. Recently, nuclear magnetic resonance spin relaxation has been instrumental in demonstrating the critical role of dynamics in enzyme function in small model systems. These studies raise questions about how dynamics affect function in larger enzymes with more complex reaction coordinates. TSase is an ideal candidate given its size, oligomeric state, cooperativity, and status as a drug target. Here, as a pre-requisite to spin relaxation studies, we present the backbone and ILV methyl resonance assignments of TSase from Escherichia coli bound to a substrate analogue and cofactor.
authors: Division of Chemical Biology and Medicinal Chemistry, UNC Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA.
authors: Paul J
authors: PJ
authors: Sapienza
authors: Andrew L
authors: AL
authors: Lee
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry*; D003067:Coenzymes / Q000378:metabolism*; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000201:enzymology*; D019906:Nuclear Magnetic Resonance, Biomolecular; D009711:Nucleotides / Q000737:chemistry*; D017433:Protein Structure, Secondary; D013940:Thymidylate Synthase / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D003067:Coenzymes; D009711:Nucleotides; D013940:Thymidylate Synthase
references: Protein Expr Purif. 2000 Jul;19(2):265-70
references: 10873540
references: Science. 2006 Sep 15;313(5793):1638-42
references: 16973882
references: J Biomol NMR. 2009 Aug;44(4):213-23
references: 19548092
references: Methods Mol Biol. 2004;278:313-52
references: 15318002
references: J Biomol NMR. 1995 Nov;6(3):277-93
references: 8520220
references: Biochemistry. 1995 Feb 7;34(5):1469-74
references: 7849005
references: Biochemistry. 1997 Apr 8;36(14):4212-22
references: 9100016
references: Biochemistry. 2003 Jan 21;42(2):239-47
references: 12525150
references: Chem Biol. 2001 Oct;8(10):981-95
references: 11590022
references: Nucleic Acids Res. 2004 Jul 1;32(Web Server issue):W500-2
references: 15215436
references: Proc Natl Acad Sci U S A. 2007 Jul 17;104(29):11981-6
references: 17615241
references: Nature. 2005 Nov 3;438(7064):117-21
references: 16267559