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J Gen Physiol. Mar 20, 1950; 33(4): 349–362.
PMCID: PMC2147187
CRYSTALLINE DESOXYRIBONUCLEASE
I. ISOLATION AND GENERAL PROPERTIES SPECTROPHOTOMETRIC METHOD FOR THE MEASUREMENT OF DESOXYRIBONUCLEASE ACTIVITY
M. Kunitz
From the Laboratories of The Rockefeller Institute for Medical Research, Princeton, New Jersey
Received October 3, 1949
Abstract
A crystalline enzyme capable of digesting thymus nucleic acid (desoxyribonucleic acid) has been isolated from fresh beef pancreas. The enzyme called "desoxyribonuclease" is a protein of the albumin type. Its molecular weight is about 60,000 and its isoelectric point is near pH 5.0. It contains about 8 per cent tyrosine and 2 per cent tryptophane. It is readily denatured by heat. The denaturation is reversible if heated in dilute acid at pH about 3.0. The digestion of thymus nucleic acid by crystalline desoxyribonuclease is accompanied by a gradual increase in the specific absorption of ultraviolet light by the acid. The spectrophotometric measurement of the rate of increase in the light absorption can be conveniently used as a general method for estimating desoxyribonuclease activity. Details are given of the method for isolation of crystalline desoxyribonuclease and of the spectrophotometric procedure for the measurement of desoxyribonuclease activity.
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