Deoxyribonucleoside kinases belonging to the thymidine kinase 2 (TK2)-like group vary significantly in substrate specificity, kinetics and feed-back regulation
(2002) In Journal of Molecular Biology 315(4). p.529-540- Abstract
In eukaryotic cells deoxyribonucleoside kinases belonging to three phylogenetic sub-families have been found: (i) thymidine kinase 1 (TK1)-like enzymes, which are strictly pyrimidine deoxyribonucleoside-specific kinases; (ii) TK2-like enzymes, which include pyrimidine deoxyribonucleoside kinases and a single multisubstrate kinase from Drosophila melanogaster (Dm-dNK); and (iii) deoxycytidine/deoxyguanosine kinase (dCK/dGK)-like enzymes, which are deoxycytidine and/or purine deoxyribonucleoside-specific kinases. We cloned and characterized two new deoxyribonucleoside kinases belonging to the TK2-like group from the insect Bombyx mori and the amphibian Xenopus laevis. The deoxyribonucleoside kinase from B. mori (Bm-dNK) turned out to be a... (More)
In eukaryotic cells deoxyribonucleoside kinases belonging to three phylogenetic sub-families have been found: (i) thymidine kinase 1 (TK1)-like enzymes, which are strictly pyrimidine deoxyribonucleoside-specific kinases; (ii) TK2-like enzymes, which include pyrimidine deoxyribonucleoside kinases and a single multisubstrate kinase from Drosophila melanogaster (Dm-dNK); and (iii) deoxycytidine/deoxyguanosine kinase (dCK/dGK)-like enzymes, which are deoxycytidine and/or purine deoxyribonucleoside-specific kinases. We cloned and characterized two new deoxyribonucleoside kinases belonging to the TK2-like group from the insect Bombyx mori and the amphibian Xenopus laevis. The deoxyribonucleoside kinase from B. mori (Bm-dNK) turned out to be a multisubstrate kinase like Dm-dNK. But uniquely for a deoxyribonucleoside kinase, Bm-dNK displayed positive cooperativity with all four natural deoxyribonucleoside substrates. The deoxyribonucleoside kinase from X. laevis (Xen-PyK) resembled closely the human and mouse TK2 enzymes displaying their characteristic Michaelis-Menten kinetic with deoxycytidine and negative cooperativity with its second natural substrate thymidine. Bm-dNK, Dm-dNK and Xen-PyK were shown to be homodimers. Significant differences in the feedback inhibition by deoxyribonucleoside triphosphates between these three enzymes were found. The insect multisubstrate deoxyribonucleoside kinases Bm-dNK and Dm-dNK were only inhibited by thymidine triphosphate, while Xen-PyK was inhibited by thymidine and deoxycytidine triphosphate in a complex pattern depending on the deoxyribonucleoside substrate. The broad substrate specificity and different feedback regulation of the multisubstrate insect deoxyribonucleoside kinases may indicate that these enzymes have a different functional role than the other members of the TK2-like group.
(Less)
- author
- Knecht, Wolfgang LU ; Petersen, Gitte Ebert ; Munch-Petersen, Birgitte LU and Piskur, Jure LU
- publishing date
- 2002-01-25
- type
- Contribution to journal
- publication status
- published
- keywords
- Amino Acid Sequence, Animals, Bombyx/enzymology, Chickens, Chromatography, Gel, Databases, Genetic, Deoxycytidine/metabolism, Drosophila melanogaster/enzymology, Feedback, Physiological, Humans, Inhibitory Concentration 50, Kinetics, Mice, Models, Biological, Models, Molecular, Molecular Sequence Data, Phosphotransferases (Alcohol Group Acceptor)/antagonists & inhibitors, Phylogeny, Protein Structure, Quaternary, Recombinant Fusion Proteins/antagonists & inhibitors, Sequence Alignment, Sequence Deletion/genetics, Substrate Specificity, Thymidine Kinase/chemistry, Xenopus laevis/genetics
- in
- Journal of Molecular Biology
- volume
- 315
- issue
- 4
- pages
- 529 - 540
- publisher
- Elsevier
- external identifiers
-
- pmid:11812127
- scopus:0036306450
- ISSN
- 0022-2836
- DOI
- 10.1006/jmbi.2001.5257
- language
- English
- LU publication?
- no
- id
- e75cddd2-6a5d-4305-9f78-e500b31d177b
- date added to LUP
- 2020-07-22 14:25:16
- date last changed
- 2024-01-02 15:00:48
@article{e75cddd2-6a5d-4305-9f78-e500b31d177b, abstract = {{<p>In eukaryotic cells deoxyribonucleoside kinases belonging to three phylogenetic sub-families have been found: (i) thymidine kinase 1 (TK1)-like enzymes, which are strictly pyrimidine deoxyribonucleoside-specific kinases; (ii) TK2-like enzymes, which include pyrimidine deoxyribonucleoside kinases and a single multisubstrate kinase from Drosophila melanogaster (Dm-dNK); and (iii) deoxycytidine/deoxyguanosine kinase (dCK/dGK)-like enzymes, which are deoxycytidine and/or purine deoxyribonucleoside-specific kinases. We cloned and characterized two new deoxyribonucleoside kinases belonging to the TK2-like group from the insect Bombyx mori and the amphibian Xenopus laevis. The deoxyribonucleoside kinase from B. mori (Bm-dNK) turned out to be a multisubstrate kinase like Dm-dNK. But uniquely for a deoxyribonucleoside kinase, Bm-dNK displayed positive cooperativity with all four natural deoxyribonucleoside substrates. The deoxyribonucleoside kinase from X. laevis (Xen-PyK) resembled closely the human and mouse TK2 enzymes displaying their characteristic Michaelis-Menten kinetic with deoxycytidine and negative cooperativity with its second natural substrate thymidine. Bm-dNK, Dm-dNK and Xen-PyK were shown to be homodimers. Significant differences in the feedback inhibition by deoxyribonucleoside triphosphates between these three enzymes were found. The insect multisubstrate deoxyribonucleoside kinases Bm-dNK and Dm-dNK were only inhibited by thymidine triphosphate, while Xen-PyK was inhibited by thymidine and deoxycytidine triphosphate in a complex pattern depending on the deoxyribonucleoside substrate. The broad substrate specificity and different feedback regulation of the multisubstrate insect deoxyribonucleoside kinases may indicate that these enzymes have a different functional role than the other members of the TK2-like group.</p>}}, author = {{Knecht, Wolfgang and Petersen, Gitte Ebert and Munch-Petersen, Birgitte and Piskur, Jure}}, issn = {{0022-2836}}, keywords = {{Amino Acid Sequence; Animals; Bombyx/enzymology; Chickens; Chromatography, Gel; Databases, Genetic; Deoxycytidine/metabolism; Drosophila melanogaster/enzymology; Feedback, Physiological; Humans; Inhibitory Concentration 50; Kinetics; Mice; Models, Biological; Models, Molecular; Molecular Sequence Data; Phosphotransferases (Alcohol Group Acceptor)/antagonists & inhibitors; Phylogeny; Protein Structure, Quaternary; Recombinant Fusion Proteins/antagonists & inhibitors; Sequence Alignment; Sequence Deletion/genetics; Substrate Specificity; Thymidine Kinase/chemistry; Xenopus laevis/genetics}}, language = {{eng}}, month = {{01}}, number = {{4}}, pages = {{529--540}}, publisher = {{Elsevier}}, series = {{Journal of Molecular Biology}}, title = {{Deoxyribonucleoside kinases belonging to the thymidine kinase 2 (TK2)-like group vary significantly in substrate specificity, kinetics and feed-back regulation}}, url = {{http://dx.doi.org/10.1006/jmbi.2001.5257}}, doi = {{10.1006/jmbi.2001.5257}}, volume = {{315}}, year = {{2002}}, }