Enzymic degradation of deoxyribonucleic acid I. Purification of acid deoxyribonuclease

Frédéricq, E.; Oth, A.

doi:10.1016/0006-3002(58)90186-0

Cited by 27 publications

(9 citation statements)

References 9 publications

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“…Nevertheless, it can be concentrated without difficulty. The consistent increase in total activity observed during the first two steps of the fractionation process has also been noted with other tissues (8,11,14), and m a y well be due to removal of inhibitors. The crystals from step 4 have been termed "inert," since they are not DNase.…”

Section: Discussionsupporting

confidence: 78%

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Deoxyribonuclease from Salmon Testes

McDonald

1962

The Journal of General Physiology

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A procedure is described for the purification of salmon testis deoxyribonuclease II b y means of acid extraction, fractional precipitation with ammonium sulfate, heat denaturation of extraneous proteins, and ethanol fractionation. This process separates the deoxyribonuclease activity from that of ribonuclease, phosphatase, phosphodiesterase, and protease. Over 50 per cent of the activity is retained with an over-all enrichment of 20,000-fold. The enzyme degrades both nadve and heat-denatured DNA, but the rate of degradation of the latter is only one-tenth that of the former. It does not hydrolyze apurinic acid.The enzyme is most stable in the pH range 4 to 5. Electrolytes are essential for the expression of its activity: monovalent ions satisfy the requirement, but divalent ones are much more effective. Above a certain optimum concentration, each electrolyte is inhibitory. The pH of maximal activity, under conditions of optimal ionic strength, is 4.8; the temperature optimum is near to 55°C.At least two types of deoxyribonuclease (DNase) have been identified in mammalian tissues. The best known is the by now "classical" crystalline DNase from beef pancreas (1, 2), representative of the DNase I (or neutral) class. This type is active in neutral solution and requires magnesium or other divalent cations (1-4) for expression of activity. The DNase II (or acid) class is most active at p H 4 to 5, and its electrolyte requirement can be satisfied with either mono-or divalent cations (5-8). DNase II predominates in most mammalian tissues (9).Procedures have been developed for the purification of DNase II from spleen and thymus (10-15). None of them appears to be operationally satisfactory for routine preparation of large amounts of enzyme. Furthermore, those purified DNases that have been tested still show measurable traces of ribonuclease and phosphodiesterase. Since the DNase II content of normal tissues appears to be intimately related to their capacity for synthesis of D N A 77

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Section: Discussionsupporting

confidence: 78%

“…Our estimates indicate that the salmon testis DNase is at least ten times as potent as the best preparations from thymus and spleen (10)(11)(12)(13)(14)(15).…”

Section: Discussionmentioning

confidence: 90%

Deoxyribonuclease from Salmon Testes

McDonald

1962

The Journal of General Physiology

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“…Chromatographic Purification.-After many smallscale trials of chromatographic purification on substances already used by previous authors, namely, Amberlite IRC-50 (Koszalka ei al., 1957), CMand DEAE-cellulose (Shimomura and Laskowski, 1957;Maver et al, 1959;Maver and Greco, 1962; Hodes and Swenson, 1962), hydroxyapatite (Fredericq and Oth, 1958), and combinations of those methods, two procedures (A and B, vide infra) were worked out.…”

Section: Isolation Of the Enzymementioning

confidence: 99%

Studies on Acid Deoxyribonuclease. II. Isolation and Characterization of Spleen-Acid Deoxyribonuclease^*

Bernardi¹,

Griffe²

1964

Biochemistry

102

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“…Several methods for the partial purification of the enzyme had been proposed (45)(46)(47)(48)(49)(50)(51)(52)(53) and its ionic requirements had been investigated to some extent (54430). The terminal oligonucleotides and the specificity of the enzyme had been studied (54) and it had been shown that all fragments were terminated in 3'-phosphates (54).…”

Section: Oiorqio Bernard1mentioning

confidence: 99%

Mechanism of Action and Structure of Acid Deoxyribonuclease

Bernardi¹

1968

Advances in Enzymology - And Related Areas of Molecular Biology

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Enzymic degradation of deoxyribonucleic acid I. Purification of acid deoxyribonuclease

Cited by 27 publications

References 9 publications

Deoxyribonuclease from Salmon Testes

Deoxyribonuclease from Salmon Testes

Studies on Acid Deoxyribonuclease. II. Isolation and Characterization of Spleen-Acid Deoxyribonuclease^*

Mechanism of Action and Structure of Acid Deoxyribonuclease

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Enzymic degradation of deoxyribonucleic acid I. Purification of acid deoxyribonuclease

Cited by 27 publications

References 9 publications

Deoxyribonuclease from Salmon Testes

Deoxyribonuclease from Salmon Testes

Studies on Acid Deoxyribonuclease. II. Isolation and Characterization of Spleen-Acid Deoxyribonuclease*

Mechanism of Action and Structure of Acid Deoxyribonuclease

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Product

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Studies on Acid Deoxyribonuclease. II. Isolation and Characterization of Spleen-Acid Deoxyribonuclease^*