Author/Editor     Knecht, Wolfang; Petersen, Gitte Ebert; Sandrini, Michael P; Sondergaard, Leif; Munch-Petersen, Birgitte; Piškur, Jure
Title     Mosquito has a single multisubstrate deoxyribonucleoside kinase characterized by unique substrate specificity
Type     članek
Source     Nucleic Acids Res purification/*metabolism
Vol. and No.     Letnik 31, št. 6
Publication year     2003
Volume     str. 1665-72
Language     eng
Abstract     In mammals four deoxyribonucleoside kinases, with a relatively restricted specificity, catalyze the phosphorylation of the four natural deoxyribonucleosides. When cultured mosquito cells, originating from the malaria vector Anopheles gambiae, were examined for deoxyribonucleoside kinase activities, only a single enzyme was isolated. Subsequently, the corresponding gene was cloned and over-expressed. While the mosquito kinase (Ag-dNK) phosphorylated all four natural deoxyribonucleosides, it displayed an unexpectedly higher relative efficiency for the phosphorylation of purine versus pyrimidine deoxyribonucleosides than the fruit fly multisubstrate deoxyribonucleoside kinase (EC 2.7.1.145). In addition, Ag-dNK could also phosphorylate some medically interesting nucleoside analogs, like stavudine (D4T), 2-chloro-deoxyadenosine (CdA) and 5-bromo-vinyl-deoxyuridine (BVDU). Although the biological significance of multisubstrate deoxyribonucleoside kinases and their diversity among insects remains unclear, the observed variation provides a whole range of applications, as species specific and highly selective targets for insecticides, they have a potential to be used in the enzymatic production of various (di-)(deoxy-)ribonucleoside monophosphates, and as suicide genes in gene therapy.
Descriptors     AMINO ACID SEQUENCE
ANIMALS
ANOPHELES
CELL LINE
CHROMATOGRAPHY, DEAE-CELLULOSE
CLONING, MOLECULAR
DNA, COMPLEMENTARY
ELECTROPHORESIS, POLYACRYLAMIDE GEL
KINETICS
MOLECULAR SEQUENCE DATA
PHOSPHORYLATION
PHOSPHOTRANSFERASES (ALCOHOL GROUP ACCEPTOR)
RECOMBINANT PROTEINS
SEQUENCE ANALYSIS, DNA
SEQUENCE HOMOLOGY
SUBSTRATE SPECIFICITY