Two pH optima of adenosine 5'-triphosphate dependent deoxyribonuclease from Bacillus laterosporus.
The various catalytic activities of the ATP-dependent deoxyribonuclease (DNase) of Bacillus laterosporus have pH optima at 6.3 and 8.3. Although the pH profile of ATP-dependent DNase activity on duplex DNA is bell shaped with a maximum at about pH 8.3, ATP-dependent DNAse activity on single-stranded DNA has optima at pH 6.3 and 8.3. ATPase activities dependent on double-stranded and single-stranded DNA have a high bell-shaped peak with a maximum at pH 6.3 with a low and broad shoulder at about pH 8.3. ATP-independent DNase activity also has optima at pH 6.3 and 8.3. The ratio of the amount of ATP hydrolyzed per number of cleaved phosphodiester bonds in DNA increases with decrease in the pH value of the reaction. The ratios obtained at pH 8.3 and 6.3 were respectively about 3 and 22 with duplex DNA as substrate and 5 and 17 with single-stranded DNA as substrate. Formation of a single-stranded region of 15000-20000 nucleotides, which is linked to duplex DNA and about half of which has 3'-hydroxyl termini, was observed at about pH 6.3, but not at above pH 7.5. Furthermore, the optimum concentrations of divalent cations for the activity producing the single-stranded region and the activity hydrolyzing ATP were identical (3 mM Mn2+ or 5 mM Mg2+). Thus the two activities are closely related. These results indicate that the enzyme has two different modes of action on duplex DNA which are modulated by the pH.
D000255:Adenosine Triphosphate / Q000494:pharmacology*; D001407:Bacillus / Q000201:enzymology*; D004277:DNA, Single-Stranded / Q000378:metabolism; D004279:DNA, Viral / Q000378:metabolism; D003851:Deoxyribonucleases / Q000378:metabolism*; D004720:Endonucleases; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D015719:Single-Strand Specific DNA and RNA Endonucleases; D013604:T-Phages
D004277:DNA, Single-Stranded; D004279:DNA, Viral; D000255:Adenosine Triphosphate; D003851:Deoxyribonucleases; D004720:Endonucleases; D015719:Single-Strand Specific DNA and RNA Endonucleases