Open AccessCloning, expression, purification, crystallization and preliminary X‐ray crystallographic study of thymidylate kinase (TTHA1607) from Thermus thermophilus HB8
Author(s) -
Chaudhary Santosh Kumar,
Jeyakanthan Jeyaraman,
Sekar Kanagaraj
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309112050208
Subject(s) - thermus thermophilus , thymidylate synthase , enzyme , dimer , orthorhombic crystal system , kinase , crystallography , biology , stereochemistry , microbiology and biotechnology , chemistry , biochemistry , crystal structure , escherichia coli , gene , genetics , cancer , fluorouracil , organic chemistry
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 P 2 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 .