Characterization of oligomeric and kinetic properties of tomato thymidine kinase 1

Publikation: Bidrag til tidsskriftTidsskriftartikelfagfællebedømt

  • Zeeshan Mutahir
  • Nicolai Balle Larsen
  • Louise Slot Christiansen
  • Karl Magnus Andersson
  • Ricardo Rico
  • Sofia Mebrahtu Wisen
  • Anders Ranegaard Clausen
  • Birgitte Munch-Petersen
  • Jure Piškur

The gene encoding thymidine kinase 1 from tomato (to TK1) has in combination with azidothymidine (AZT) recently been proposed as a powerful suicide gene for anticancer gene therapy. The to TK1/AZT combination has been demonstrated to have several advantages for the treatment of glioblastomas because AZT can easily penetrate the blood-brain barrier and to TK1 can efficiently phosphorylate AZT and also AZT-monophosphate. In a pursuit to further understand the properties of to TK1, we examined the oligomerization properties of recombinant to TK1 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 to TK1. For human TK1 pretreated with ATP, the Km value decreased 20-fold, but to TK1's Km value did not show a dependence on the presence or absence of ATP. Furthermore, to TK1 was always found in a highly active form.

OriginalsprogEngelsk
TidsskriftNucleosides, Nucleotides and Nucleic Acids
Vol/bind30
Udgave nummer12
Sider (fra-til)1223-1226
Antal sider4
ISSN1525-7770
DOI
StatusUdgivet - 2011
Eksternt udgivetJa

Bibliografisk note

(Ekstern)

ID: 255884357