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

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

SLU Authors

• Wang, Liya

  • Department of Animal Biosciences, Swedish University of Agricultural Sciences
    

• Eriksson, Staffan

  • Department of Animal Biosciences, Swedish University of Agricultural Sciences
    

UKÄ Subject classification

Animal and Dairy Science
Veterinary Science

Publication identifier

• DOI: https://doi.org/10.1080/15257770802146510

Permanent link to this page (URI)

https://res.slu.se/id/publ/20796