Bromovinyl-deoxyuridine: A selective substrate for mitochondrial thymidine kinase in cell extracts.

Biochemical and Biophysical Research Communications
Elisa FranzolinVera Bianchi

Abstract

Cellular models of mitochondrial thymidine kinase (TK2) deficiency require a reliable method to measure TK2 activity in whole cell extracts containing two interfering deoxyribonucleoside kinases, thymidine kinase 1 (TK1) and deoxycytidine kinase. We tested the value of the thymidine analog (E)-5-(2-bromovinyl)-2'-deoxyuridine (BVDU) as a TK2-specific substrate. With extracts of OSTTK1- cells containing TK2 as the only thymidine kinase and a highly specific TK2 inhibitor we established conditions to detect the low TK2 activity commonly present in cells. With extracts of TK1-proficient osteosarcoma cells and normal human fibroblasts we showed that BVDU, but not 1-(beta-d-arabinofuranosyl)thymine (Ara-T), discriminates TK2 activity even in the presence of 100-fold excess TK1. A comparison with current procedures based on TK2 inhibition demonstrated the better performance of the new TK2 assay. When cultured human fibroblasts passed from proliferation to quiescence TK2 activity increased by 3-fold, stressing the importance of TK2 function in the absence of TK1.

References

Oct 4, 1977·Biochimica Et Biophysica Acta·B Munch-Petersen, G Tyrsted
Jan 1, 1995·Pharmacology & Therapeutics·E S Arnér, S Eriksson
Mar 28, 1997·The Journal of Biological Chemistry·M Johansson, A Karlsson
Aug 14, 1999·The Journal of Biological Chemistry·M JohanssonA Karlsson
Sep 6, 2002·Journal of Medicinal Chemistry·Ana-Isabel HernándezMaría-Jesús Pérez-Pérez
Oct 2, 2003·Proceedings of the National Academy of Sciences of the United States of America·Giovanna PontarinVera Bianchi
Jan 30, 2004·The Journal of Biological Chemistry·Chiara RampazzoVera Bianchi
Jan 27, 2006·The Journal of Biological Chemistry·Pelle HåkanssonLars Thelander

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Citations

Feb 13, 2008·European Journal of Nuclear Medicine and Molecular Imaging·Omid S TehraniAnthony F Shields
Jan 9, 2013·The Journal of Biological Chemistry·Miriam FranginiChiara Rampazzo
Apr 15, 2010·Nucleosides, Nucleotides & Nucleic Acids·Mia BjerkeAnna Karlsson
Apr 25, 2008·Human Molecular Genetics·Xiaoshan ZhouAnna Karlsson
Jul 24, 2012·Journal of Biochemistry·Tadanobu TakahashiTakashi Suzuki
Jun 22, 2010·Mutation Research·Chiara RampazzoVera Bianchi
May 7, 2008·Medicinal Research Reviews·María-Jesús Pérez-PérezJan Balzarini
Feb 12, 2017·Cell Death and Differentiation·Deborah PajalungaMarco Crescenzi
Apr 18, 2008·The Journal of Biological Chemistry·Luigi LeanzaVera Bianchi
Sep 15, 2007·Molecular Pharmacology·Nicola SolaroliAnna Karlsson
Aug 3, 2021·Annals of Neurology·Carlos Lopez-GomezMichio Hirano
Dec 22, 2006·Journal of Medicinal Chemistry·Ana-Isabel HernandezMaría-Jesús Pérez-Pérez

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