Cloning and expression of human mitochondrial deoxyguanosine kinase cDNA

FEBS Letters

Hellman, Ulf¹  Eriksson, Staffan²  Wang, Liya² 

[1] The Ludwig Institute of Cancer Research, The Biomedical Center, Uppsala, Sweden;Department of Veterinary Medical Chemistry, Swedish University of Agricultural Sciences, The Biomedical Center, Box 575, S-751 23 Uppsala, Sweden

关键词: Mitochondria;    DNA precursor synthesis;    Deoxynucleoside kinase;    Sequence;    Conserved motif;    Chemotherapy;   

DOI  :  10.1016/0014-5793(96)00623-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Mammalian mitochondrial deoxyguanosine kinase (dGK) is responsible for phosphorylation of purine deoxyribonucleosides in the mitochondrial matrix. Using a RT-PCR-generated probe, based on amino acid sequence information from proteolytic fragments of purified bovine dGK, we have cloned a cDNA from a human brain cDNA library that encodes a 30 kDa protein. The deduced amino acid sequence of this protein included the sequence of all six peptides isolated and sequenced from purified dGK. Expression and purification of recombinant protein from induced Escherichia coli extracts revealed that it catalyses efficient phosphorylation of dGuo, arabinosyl guanine, dAdo, 2-chloro-2′-deoxyadenosine and dIno similar to purified dGK. Northern blot analysis demonstrated one dominant positive mRNA of 1.35 kb and it was found in several tissues at similar levels. The coding sequence of dGK showed 46% identity to the coding sequence of cytosolic deoxycytidine kinase, and conserved sequence motifs among the known deoxynucleoside kinase were identified.

【 授权许可】

Unknown