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 |
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Authors: | Wang Liya Eriksson Staffan |
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Affiliation: | Department of Anatomy, Physiology and Biochemistry, Section of Veterinary Medical Biochemistry, Swedish University of Agricultural Sciences, The Biomedical Centre, Uppsala, Sweden. |
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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 micro 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 micro M but was 500 at 2.5 micro M. Therefore, for reliable measurement of TK2 activity higher BvdU concentration should be used. |
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