Purification and properties of ribonuclease H of calf thymus.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 70 ·No. 7 ·1973-07-00 ·Pages 1959-63

Stavrianopoulos JG, Chargaff E

Abstract

Ribonuclease H of calf thymus has been purified better than 3000-fold to yield an almost homogeneous preparation. The enzyme, which comprises about 0.03% of the total protein in the initial extract, is a slightly acidic protein (pI = 4.95) of molecular weight of about 64,000, possibly composed of subunits. The enzyme requires a metal ion for activation; the conditions for activation by Mg, Co, and Mn are described. It is inhibited by S-adenosylmethionine. The substrates cleaved were poly(dT).poly(rA) and the DNA-RNA hybrid made from phage f1 DNA; ribosomal RNA was not attacked.

MeSH Terms

Animals Cattle Chromatography, DEAE-Cellulose Chromatography, Gel Cobalt/pharmacology DNA, Viral Electrophoresis, Polyacrylamide Gel Enzyme Activation/drug effects Magnesium/pharmacology Manganese/pharmacology Nucleic Acid Hybridization Polynucleotides/metabolism RNA Ribonucleases/antagonists & inhibitors,isolation & purification,metabolism S-Adenosylmethionine/pharmacology Thymus Gland/enzymology

Chemicals

DNA, Viral Polynucleotides Cobalt Manganese RNA S-Adenosylmethionine Ribonucleases Magnesium

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Stavrianopoulos J G

Chargaff E

References (12)

12 references, click to expand

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Proc Natl Acad Sci U S A. 1971 Sep;68(9):2207-11 PMID: 4943793
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Article Info

Journal

Proceedings of the National Academy of Sciences of the United States of America

Abbr.

Proc Natl Acad Sci U S A

ISSN

0027-8424

DOI

10.1073/pnas.70.7.1959

Published

1973-07-00

Pages

1959-63

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC433642

Subset

External Links

PubMed source DOI: 10.1073/pnas.70.7.1959 PMC Full Text View journal details

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Purification and properties of ribonuclease H of calf thymus.

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