title: Expression of tomato thymidine kinase 1 by means of the baculovirus expression vector system.
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 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.
authors.affiliation: a Department of Biology and Lund Protein Production Platform , Lund University , Lund , Sweden.
authors.forename: Juan Manuel
authors.initials: JM
authors.lastname: Orozco Rodriguez
authors.affiliation: a Department of Biology and Lund Protein Production Platform , Lund University , Lund , Sweden.
authors.forename: Mohamad
authors.initials: M
authors.lastname: Nesrini
authors.affiliation: a Department of Biology and Lund Protein Production Platform , Lund University , Lund , Sweden.
authors.forename: Louise Slot
authors.initials: LS
authors.lastname: Christiansen
authors.affiliation: a Department of Biology and Lund Protein Production Platform , Lund University , Lund , Sweden.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Knecht
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D018551:Solanum lycopersicum / Q000201:enzymology*; D010940:Plant Proteins / Q000096:biosynthesis / Q000235:genetics*; D061987:Sf9 Cells; D018411:Spodoptera; D013937:Thymidine Kinase / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D010940:Plant Proteins; D013937:Thymidine Kinase; C092717:thymidine kinase 1
keywords: Nucleoside salvage pathway; baculovirus expression vector system; nucleotide metabolism; protein production; suicide gene therapy