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Expression of tomato thymidine kinase 1 by means of the baculovirus expression vector system

Orozco Rodriguez, Juan Manuel; Nesrini, Mohamad; Christiansen, Louise Slot LU and Knecht, Wolfgang LU (2016) In Nucleosides, Nucleotides and Nucleic Acids 35(10-12). p.691-698
Abstract

Tomato thymidine kinase 1 (ToTK1) is a deoxyribonucleoside kinase (dNK) that has been subject to study because of its potential to phosphorylate the nucleoside analogue 3-azido-2,3-dideoxythymidine (azidothymidine, AZT) equally well as its natural substrate thymidine (dThd). The combination of ToTK1 and AZT has been tested in two animal studies for its efficiency and use in suicide gene therapy for malignant glioma. The determination of the 3D structure of ToTK1 might shed light on the structure–function relationships of nucleoside activation by this enzyme and thereby show routes toward further improvement of ToTK1 and other TK1-like dNKs for suicide gene therapy. Here we report the successful expression of both full-length ToTK1 and a... (More)

Tomato thymidine kinase 1 (ToTK1) is a deoxyribonucleoside kinase (dNK) that has been subject to study because of its potential to phosphorylate the nucleoside analogue 3-azido-2,3-dideoxythymidine (azidothymidine, AZT) equally well as its natural substrate thymidine (dThd). The combination of ToTK1 and AZT has been tested in two animal studies for its efficiency and use in suicide gene therapy for malignant glioma. The determination of the 3D structure of ToTK1 might shed light on the structure–function relationships of nucleoside activation by this enzyme and thereby show routes toward further improvement of ToTK1 and other TK1-like dNKs for suicide gene therapy. Here we report the successful expression of both full-length ToTK1 and a C-terminal truncated ToTK1 in Spodoptera frugiperda and Trichoplusia ni insect cells using the baculovirus expression vector system. This constitutes a further step on the road to determine the 3D structure of the first TK1 of plant origin, but also an enzyme with great potential for dNK-mediated suicide gene therapy.

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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
baculovirus expression vector system, Nucleoside salvage pathway, nucleotide metabolism, protein production, suicide gene therapy
in
Nucleosides, Nucleotides and Nucleic Acids
volume
35
issue
10-12
pages
8 pages
publisher
Taylor & Francis
external identifiers
  • scopus:85000838074
  • wos:000389143100023
ISSN
1525-7770
DOI
10.1080/15257770.2016.1139126
language
English
LU publication?
yes
id
a90d1681-5ff5-4a21-b819-c9f991972579
date added to LUP
2016-12-16 12:42:03
date last changed
2017-09-18 11:31:39
@article{a90d1681-5ff5-4a21-b819-c9f991972579,
  abstract     = {<p>Tomato thymidine kinase 1 (ToTK1) is a deoxyribonucleoside kinase (dNK) that has been subject to study because of its potential to phosphorylate the nucleoside analogue 3-azido-2,3-dideoxythymidine (azidothymidine, AZT) equally well as its natural substrate thymidine (dThd). The combination of ToTK1 and AZT has been tested in two animal studies for its efficiency and use in suicide gene therapy for malignant glioma. The determination of the 3D structure of ToTK1 might shed light on the structure–function relationships of nucleoside activation by this enzyme and thereby show routes toward further improvement of ToTK1 and other TK1-like dNKs for suicide gene therapy. Here we report the successful expression of both full-length ToTK1 and a C-terminal truncated ToTK1 in Spodoptera frugiperda and Trichoplusia ni insect cells using the baculovirus expression vector system. This constitutes a further step on the road to determine the 3D structure of the first TK1 of plant origin, but also an enzyme with great potential for dNK-mediated suicide gene therapy.</p>},
  author       = {Orozco Rodriguez, Juan Manuel and Nesrini, Mohamad and Christiansen, Louise Slot and Knecht, Wolfgang},
  issn         = {1525-7770},
  keyword      = {baculovirus expression vector system,Nucleoside salvage pathway,nucleotide metabolism,protein production,suicide gene therapy},
  language     = {eng},
  month        = {12},
  number       = {10-12},
  pages        = {691--698},
  publisher    = {Taylor & Francis},
  series       = {Nucleosides, Nucleotides and Nucleic Acids},
  title        = {Expression of tomato thymidine kinase 1 by means of the baculovirus expression vector system},
  url          = {http://dx.doi.org/10.1080/15257770.2016.1139126},
  volume       = {35},
  year         = {2016},
}