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Title: Micrococcus luteus correndonucleases. I. Resolution and purification of two endonucleases specific for DNA containing pyrimidine dimers. [Uv 500 J/m/sup 2/; x-radiation of 300 Krad; micrococcus lutea]

Journal Article · · J. Biol. Chem.; (United States)
OSTI ID:5112004

Five peaks of endonuclease activity showing a preference for ultraviolet-damaged DNA have been chromatographically identified from extracts of Micrococcus luteus. They are numerically designated as I to V in order of their elution from phosphocellulose (Whatman P-11) columns. The first two of these peaks have been highly purified by a combination of gel filtration and affinity chromatography and are catalytically homogeneous judging from their effect on transforming DNAs. Peak I, which has an isoelectric point of 4.7, is heat-stable, requires high ionic strength for optimal activity, acts with equal facility on ultraviolet-irradiated native and denatured DNA, and has been designated as Py--Py correndonuclease I. Peak II which has a pI value of 8.7, is heat-labile, is inhibited by high ionic strength, acts on ultraviolet-irradiated native but not denatured DNA, and has been designated as Py--Py correndonuclease II. Both enzymes are inhibited by Ca/sup 2 +/ and Zn/sup 2 +/, do not show any cofactor or sulfhydryl requirement, act optimally between pH 7.0 and 7.4, and have molecular weights between 11,000 and 15,000. Py--Py correndonuclease I requires a dose about 1.6 times that for Py--Py correndonuclease II for incision saturation of irradiated phiX174 RFI DNA.

Research Organization:
Johns Hopkins Univ., Baltimore
OSTI ID:
5112004
Journal Information:
J. Biol. Chem.; (United States), Vol. 252:18
Country of Publication:
United States
Language:
English