Expression of tomato thymidine kinase 1 by means of the baculovirus expression vector system
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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; Knecht, Wolfgang.
I: Nucleosides, Nucleotides and Nucleic Acids, Bind 35, Nr. 10-12, 2016, s. 691-698.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - Expression of tomato thymidine kinase 1 by means of the baculovirus expression vector system
AU - Orozco Rodriguez, Juan Manuel
AU - Nesrini, Mohamad
AU - Christiansen, Louise Slot
AU - Knecht, Wolfgang
N1 - (Ekstern)
PY - 2016
Y1 - 2016
N2 - 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.
AB - 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.
KW - Baculovirus expression vector system
KW - Nucleoside salvage pathway
KW - Nucleotide metabolism
KW - Protein production
KW - Suicide gene therapy
UR - http://www.scopus.com/inward/record.url?scp=85000838074&partnerID=8YFLogxK
U2 - 10.1080/15257770.2016.1139126
DO - 10.1080/15257770.2016.1139126
M3 - Journal article
C2 - 27906616
AN - SCOPUS:85000838074
VL - 35
SP - 691
EP - 698
JO - Nucleosides, Nucleotides and Nucleic Acids
JF - Nucleosides, Nucleotides and Nucleic Acids
SN - 1525-7770
IS - 10-12
ER -
ID: 255883336