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Research article2008Peer reviewed

5-bromovinyl 2'-deoxyuridine phosphorylation by mitochondrial and cytosolic thymidine kinase (TK2 and TK1) and its use in selective measurement of TK2 activity in crude extracts

Wang, Liya; Eriksson, Staffan

Abstract

Mitochondrial thymidine kinase (TK2) is responsible for phosphorylation of thymidine and deoxycytidine and plays a crucial role in mitochondrial DNA precursor synthesis. TK2 is expressed in all tissues at low levels complicating accurate determinations, especially in tissues with high cytosolic thymidine kinase (TK1) activity. Recently, 5-bromovinyl 2'-deoxyuridine (BvdU) at 0.2 mu M was used to measure TK2 activity selectively. BvdU phosphorylation by pure human TK2 and TK1 was tested here, and the ratio of BvdU phosphorylation by TK2/TK1 was 91 at 0.2 mu M but was 500 at 2.5 mu M. Therefore, for reliable measurement of TK2 activity higher BvdU concentration should be used.

Keywords

thymidine kinase 2; thymidine kinase 1; activity determination; bromovinyl deoxyurindine

Published in

Nucleosides, Nucleotides and Nucleic Acids
2008, Volume: 27, number: 6-7, pages: 858-862 Publisher: TAYLOR & FRANCIS INC