Properties of an ATP-dependent deoxyribonuclease from Micrococcus luteus Evidence for a stable DNA-enzyme complex
article
The mechanism of action of the ATP-dependent DNAase from Micrococcus luteus, an enzyme which presumably is involved in the process of genetic recombination, has been investigated with partially purified or extensively purified enzyme (approximately 50 % pure). We have shown that the enzyme requires linear double-stranded DNA for activity. Double-stranded DNA with extended single-stranded regions or single-stranded DNA are degraded at a greatly reduced rate and circular double-stranded DNA is completely refractory towards the enzyme. Glycerol gradient sedimentation analysis as well as phase partition experiments have shown that the enzyme forms a stable complex with linear double-stranded DNA, but not with single-stranded DNA or circular double-stranded DNA. For this reaction ATP is not required. Maximally two enzyme molecules will bind to a linear duplex, probably one to each end. Results from kinetic experiments suggest that the formation of the enzyme-DNA complex is a necessary intermediate in the overall degradation process. Degradation after the formation of the DNA-enzyme complex is a first-order process, its rate being proportional to the concentration of the complex. At 0°C one enzyme molecule needs about 30 min to degrade one T7 DNA molecule. © 1973.
Chemicals/CAS: adenosine triphosphate, 15237-44-2, 56-65-5, 987-65-5; deoxyribonuclease, 37211-67-9; DNA polymerase, 37217-33-7; DNA, 9007-49-2; phosphorus, 7723-14-0; tritium, 10028-17-8; Adenosine Triphosphate, 56-65-5; Deoxyribonucleases, EC 3.1.-; DNA Nucleotidyltransferases, EC 2.7.7.-; DNA, 9007-49-2; DNA, Circular; DNA, Single-Stranded; Macromolecular Systems; Phosphorus Isotopes; Polysaccharides; Tritium, 10028-17-8
Chemicals/CAS: adenosine triphosphate, 15237-44-2, 56-65-5, 987-65-5; deoxyribonuclease, 37211-67-9; DNA polymerase, 37217-33-7; DNA, 9007-49-2; phosphorus, 7723-14-0; tritium, 10028-17-8; Adenosine Triphosphate, 56-65-5; Deoxyribonucleases, EC 3.1.-; DNA Nucleotidyltransferases, EC 2.7.7.-; DNA, 9007-49-2; DNA, Circular; DNA, Single-Stranded; Macromolecular Systems; Phosphorus Isotopes; Polysaccharides; Tritium, 10028-17-8
Topics
Adenosine triphosphateCircular DNADeoxyribonucleaseDNADNA polymeraseDNA, Single StrandedPhosphorusPolysaccharideSingle stranded DNATritiumBinding competitionBinding siteColiphageDensity gradient centrifugationDrug stabilityEnzymologyEscherichia coliGel chromatographyIon exchange chromatographyIsolation and purificationKineticsMacromoleculeMetabolismMicrococcusMolecular weightPolyacrylamide gel electrophoresisProtein bindingTimeAdenosine TriphosphateBinding SitesBinding, CompetitiveCentrifugation, Density GradientChromatography, DEAE-CelluloseChromatography, GelColiphagesDeoxyribonucleasesDNADNA NucleotidyltransferasesDNA, CircularDNA, Single-StrandedDrug StabilityElectrophoresis, Polyacrylamide GelEscherichia coliKineticsMacromolecular SystemsMicrococcusMolecular WeightPhosphorus IsotopesPolysaccharidesProtein BindingTime FactorsTritium
TNO Identifier
227409
ISSN
00052787
Source
BBA Section Nucleic Acids And Protein Synthesis, 299(1), pp. 65-81.
Pages
65-81
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