Inhibitory effect of pyridoxal 5'-phosphate on the DNA binding site of ATP-dependent deoxyribonuclease from Bacillus laterosporus.
Bacillus laterosporus ATP-dependent deoxyribonuclease has been found to be inhibited by pyridoxal 5'-phosphate. The inhibition is specific for pyridoxal 5'-phosphate and pyridoxal which are required in relatively high concentrations. Pyridoxamine 5'-phosphate, pyridoxamine, and pyridoxine are ineffective. The inhibition is reversed by dilution or dialysis but can be changed to an irreversible inactivation by reduction of the enzyme . pyridoxal 5'-phosphate complex with sodium borohydride. The compound is a competitive inhibitor with respect to DNA but not ATP. Moreover, the presence of DNA substrate protects the enzyme against this inactivation but the presence of ATP shows no effect. The reduced enzyme . pyridoxal 5'-phosphate complex displays a new absorption maximum at 325 nm and a fluorescence emission at 390-400 nm when excited at 325 nm which are characteristic for epsilon-N-(phosphopyridoxyl)lysine. Thus, B. laterosporus DNase appears to have an essential lysine residue at the DNA binding site of the enzyme, and the enzyme possess two different active sites, a DNA binding site and an ATP binding site.
D001407:Bacillus / Q000201:enzymology*; D001665:Binding Sites / Q000187:drug effects; D001894:Borohydrides / Q000494:pharmacology; D004247:DNA / Q000378:metabolism*; D003851:Deoxyribonucleases / Q000037:antagonists & inhibitors*; D043211:Exodeoxyribonuclease V; D005092:Exonucleases / Q000037:antagonists & inhibitors*; D011730:Pyridoxal / Q000494:pharmacology; D011732:Pyridoxal Phosphate / Q000494:pharmacology*; D011733:Pyridoxamine / Q000494:pharmacology; D013050:Spectrometry, Fluorescence
D001894:Borohydrides; D011730:Pyridoxal; D011732:Pyridoxal Phosphate; D011733:Pyridoxamine; C025364:sodium borohydride; D004247:DNA; D003851:Deoxyribonucleases; D005092:Exonucleases; D043211:Exodeoxyribonuclease V