S. Advani et Kb. Roy, Properties and secondary structure analysis of BanI endonuclease: Identification of putative active site, BIOC BIOP R, 279(1), 2000, pp. 11-16
Citations number
15
Categorie Soggetti
Biochemistry & Biophysics
Journal title
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Biochemical properties of Type II restriction enzyme BanI were characterize
d, Kinetic parameters were evaluated and an enhancement of rate was observe
d when the recognition site was located in a more central position in the s
ubstrate, suggesting that BanI locates its recognition site by a sliding me
chanism, As BanI has three cysteine residues in its primary sequence, the e
ffect of thiol inhibitors on BanI activity was also studied. Partial inhibi
tion was observed only at a very high concentration of the inhibitor indica
ting that cysteine residues are not directly involved in catalysis. The gel
electrophoretic mobility shift assay demonstrated specific complex formati
on between BanI and the DNA substrate in the presence of poly dI-dC and Mg2
+. A secondary structure analysis and comparison with EcoRI and BamHI cryst
al structure revealed a putative active site similar to that seen in BamHI
but different in the order in which the catalytic domain (central beta -she
et) and recognition domain (adjacent alpha -helix) were arranged in the pro
tein, (C) 2000 Academic Press.