A novel ADP-dependent DNA ligase from Aeropyrum pernix K1.
A gene encoding a putative ATP-dependent DNA ligase from the aerobic hyperthermophilic archaeon Aeropyrum pernix K1 was cloned and the biochemical characteristics of the resulting recombinant protein were examined. The gene (accession no. APE1094) from A. pernix encoding a 69-kDa protein showed a 39-61% identity with other ATP-dependent DNA ligases from the archaea. Normally DNA ligase is activated by NAD(+) or ATP. There has been no report about the other activators for DNA ligase. The recombinant ligase was a monomeric protein and catalyzed strand joining on a singly nicked DNA substrate in the presence of ADP and a divalent cation (Mg(2+), Mn(2+), Ca(2+) and Co(2+)) at high temperature. The optimum temperature and pH for nick-closing activity were above 70 degrees C and 7.5 degrees C, respectively. The ligase remained stable for 60 min of treatment at 100 degrees C, and the half-life was about 25 min at 110 degrees C. This is the first report of a novel hyperthermostable DNA ligase that can utilize ADP to activate the enzyme.
Jeon
Sung-Jong
SJ
The Special Division for Human Life Technology, National Institute of Advanced Industrial Science and Technology (AIST Kansai), 1-8-31 Midorigaoka, Ikeda, Osaka 563-8577, Japan.
Ishikawa
Kazuhiko
K
D000244:Adenosine Diphosphate / Q000378:metabolism*; D019843:Archaeal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D001483:Base Sequence; D002384:Catalysis; D002412:Cations / Q000378:metabolism; D003001:Cloning, Molecular; D011088:DNA Ligases / Q000235:genetics* / Q000378:metabolism*; D019620:Desulfurococcaceae / Q000201:enzymology* / Q000235:genetics; D004795:Enzyme Stability; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D013696:Temperature
D019843:Archaeal Proteins; D002412:Cations; D011994:Recombinant Proteins; D000244:Adenosine Diphosphate; D011088:DNA Ligases