Purification and properties of Mg2+-Ca2+ adenosinetriphosphatase from Escherichia coli.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 71 ·No. 7 ·1974-07-00 ·Pages 2720-4

Nelson N, Kanner BI, Gutnick DL

Abstract

A procedure for the purification of Mg(2+)-Ca(2+) adenosinetriphosphatase (EC 3.6.1.3) from E. coli, yielding relatively large amounts of highly active enzyme, is described. The enzyme consists of four nonidentical subunits. Trypsin treatment of purified enzyme yields a preparation consisting exclusively of the two larger subunits, which are sufficient for ATPase activity. Purified enzyme is inhibited by 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole; this inhibition is reversed by dithiothreitol, and the diazole is found preferentially associated with the beta-subunit of the enzyme. Antibody prepared against the trypsin-treated enzyme inhibited various ATP-dependent reactions as well as membrane-bound ATPase itself.

MeSH Terms

Adenosine Diphosphate Adenosine Triphosphatases/analysis,antagonists & inhibitors,isolation & purification Adenosine Triphosphate Antibodies Calcium Dithiothreitol Electrophoresis Escherichia coli/enzymology Magnesium Protein Binding Sodium Dodecyl Sulfate Sulfhydryl Reagents Tritium Trypsin

Chemicals

Antibodies Sulfhydryl Reagents Tritium Sodium Dodecyl Sulfate Adenosine Diphosphate Adenosine Triphosphate Trypsin Adenosine Triphosphatases Magnesium Calcium Dithiothreitol

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Nelson N

Kanner B I

Gutnick D L

References (19)

19 references, click to expand

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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.71.7.2720

Published

1974-07-00

Pages

2720-4

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC388540

Subset

External Links

PubMed source DOI: 10.1073/pnas.71.7.2720 PMC Full Text View journal details

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Purification and properties of Mg2+-Ca2+ adenosinetriphosphatase from Escherichia coli.

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