Cloning, expression, purification, crystallization and preliminary X‐ray crystallographic study of thymidylate kinase (TTHA1607) from Thermus thermophilus HB8. Issue 2 (5th February 2013)

• Record Type: Journal Article
• Authors: Chaudhary, Santosh Kumar; Jeyakanthan, Jeyaraman; Sekar, Kanagaraj
• Is Part Of: Acta crystallographica. Volume 69:Issue 2(2013:Feb.)
• Page Start: 118
• Page End: 121
• Publication Date: 2013-02-05
• Subjects: Crystallography -- Periodicals; Crystals -- Periodicals; 548
• DOI: 10.1107/S1744309112050208
• Languages: English
• ISSNs: 1744-3091

Abstract

<abstract abstract-type="main" xml:lang="en"> <title> <x xml:space="preserve">Abstract</x> </title> <p>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‐Mg²⁺ as a phosphoryl‐donor group. This enzyme is present at the junction of the <italic>de novo</italic> 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 <italic>Thermus thermophilus</italic> 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 <italic>P</italic>2₁2₁2₁, with unit‐cell parameters <italic>a</italic> = 39.50, <italic>b</italic> = 80.29, <italic>c</italic> = 122.55 Å. Preliminary studies revealed the presence of a dimer in the asymmetric unit with a Matthews coefficient (<italic>V</italic>_M) of 2.18 ų Da⁻¹.</p> </abstract>