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

Mutahir, Zeeshan LU ; Larsen, Nicolai Balle; Slot Christiansen, Louise LU ; Andersson, Karl-Magnus LU ; Rico, Ricardo; Mebrahtu Wisén, Sofia LU ; Clausen, Anders Ranegaard LU ; Munch-Petersen, Birgitte LU and Piskur, Jure LU (2011) In Nucleosides, Nucleotides & Nucleic Acids 30(12). p.1223-1226
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... (More)
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. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
ATP activation, Thymidine kinase 1, tomato (Solanum lycopersicum), oligomerization, enzyme kinetics
in
Nucleosides, Nucleotides & Nucleic Acids
volume
30
issue
12
pages
1223 - 1226
publisher
Taylor & Francis
external identifiers
  • wos:000298738500030
  • pmid:22132978
  • scopus:84855868493
ISSN
1525-7770
DOI
10.1080/15257770.2011.597629
language
English
LU publication?
yes
id
32095531-ea71-4eac-b0f8-3ce8a5620851 (old id 2274704)
date added to LUP
2012-02-06 13:48:29
date last changed
2017-01-01 03:58:29
@article{32095531-ea71-4eac-b0f8-3ce8a5620851,
  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.},
  author       = {Mutahir, Zeeshan and Larsen, Nicolai Balle and Slot Christiansen, Louise and Andersson, Karl-Magnus and Rico, Ricardo and Mebrahtu Wisén, Sofia and Clausen, Anders Ranegaard and Munch-Petersen, Birgitte and Piskur, Jure},
  issn         = {1525-7770},
  keyword      = {ATP activation,Thymidine kinase 1,tomato (Solanum lycopersicum),oligomerization,enzyme kinetics},
  language     = {eng},
  number       = {12},
  pages        = {1223--1226},
  publisher    = {Taylor & Francis},
  series       = {Nucleosides, Nucleotides & Nucleic Acids},
  title        = {Characterization of oligomeric and kinetic properties of tomato thymidine kinase 1.},
  url          = {http://dx.doi.org/10.1080/15257770.2011.597629},
  volume       = {30},
  year         = {2011},
}