title: Cloning, expression, purification, crystallization and preliminary X-ray crystallographic study of thymidylate kinase (TTHA1607) from Thermus thermophilus HB8.
abstract: Nucleotide biosynthesis plays a key role in cell survival and cell proliferation. Thymidylate kinase is an enzyme that catalyses the conversion of dTMP to dTDP using ATP-Mg(2+) as a phosphoryl-donor group. This enzyme is present at the junction of the de novo and salvage pathways; thus, any inhibitor designed against it will result in cell death. This highlights the importance of this enzyme as a drug target. Thymidylate kinase from the extremely thermophilic organism Thermus thermophilus HB8 has been expressed, purified and crystallized using the microbatch method. The crystals diffracted to a resolution of 1.83 Å and belonged to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 39.50, b = 80.29, c = 122.55 Å. Preliminary studies revealed the presence of a dimer in the asymmetric unit with a Matthews coefficient (V(M)) of 2.18 Å(3) Da(-1).
authors: Department of Physics, Indian Institute of Science, Bangalore 560 012, India.
authors: Santosh Kumar
authors: SK
authors: Chaudhary
authors: Jeyaraman
authors: J
authors: Jeyakanthan
authors: Kanagaraj
authors: K
authors: Sekar
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D008969:Molecular Sequence Data; D009703:Nucleoside-Phosphate Kinase / Q000737:chemistry* / Q000302:isolation & purification*; D016415:Sequence Alignment; D016963:Thermus thermophilus / Q000201:enzymology*
chemical_list: D009703:Nucleoside-Phosphate Kinase; C020093:dTMP kinase
keywords: Thermus thermophilus HB8; thymidylate kinase (TTHA1607)
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