Characterization of oligomeric and kinetic properties of tomato thymidine kinase 1.

Research output: Contribution to journalArticle

Abstract

The gene encoding thymidine kinase 1 from tomato (toTK1) has in combination with azidothymidine (AZT) recently been proposed as a powerful suicide gene for anticancer gene therapy. The toTK1/AZT combination has been demonstrated to have several advantages for the treatment of glioblastomas because AZT can easily penetrate the blood-brain barrier and toTK1 can efficiently phosphorylate AZT and also AZT-monophosphate. In a pursuit to further understand the properties of toTK1, we examined the oligomerization properties of recombinant toTK1 and its effect on enzyme kinetics. Previously, it has been shown that human TK1 is a dimer in the absence of ATP and a tetramer if preincubated with ATP. However, we show here that ATP preincubation did not result in a structural shift from dimer to tetramer in toTK1. For human TK1 pretreated with ATP, the K(m) value decreased 20-fold, but toTK1's K(m) value did not show a dependence on the presence or absence of ATP. Furthermore, toTK1 was always found in a highly active form.

Details

Authors
  • Zeeshan Mutahir
  • Nicolai Balle Larsen
  • Louise Slot Christiansen
  • Karl-Magnus Andersson
  • Ricardo Rico
  • Sofia Mebrahtu Wisén
  • Anders Ranegaard Clausen
  • Birgitte Munch-Petersen
  • Jure Piskur
Organisations
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Biological Sciences

Keywords

  • ATP activation, Thymidine kinase 1, tomato (Solanum lycopersicum), oligomerization, enzyme kinetics
Original languageEnglish
Pages (from-to)1223-1226
JournalNucleosides, Nucleotides & Nucleic Acids
Volume30
Issue number12
Publication statusPublished - 2011
Publication categoryResearch
Peer-reviewedYes