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PMID: 6269086 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

DNA gyrase: affinity chromatography on novobiocin-Sepharose and catalytic properties.

Nucleic acids research ·Vol. 9 ·No. 15 ·1981-08-11 ·Pages 3589-603

Staudenbauer WL, Orr E

Abstract

Novobiocin-Sepharose was prepared by coupling of novobiocin to Epoxy-activated Sepharose 6B and used as an affinity adsorbent. Four novobiocin-binding proteins were isolated from crude extracts of Escherichia coli with molecular weights of 105, 92, 85 and 40 kdal. The two larger proteins were identified as the A subunit (gyrA protein) and the B subunit (gyrB protein) of DNA gyrase topoisomerase II). By this method the two gyrase components can be easily separated and purified in high yield. Although both proteins are involved in the ATP-dependent supercoiling of relaxed plasmid DNA, only the gyrB protein is required for catalyzing the cleavage of ATP. The gyrB protein ATPase activity is competitively inhibited by novobiocin and related coumarin antibiotics. ATP hydrolysis is unaffected by the addition of either gyrA protein or DNA but stimulated in the presence of both.

MeSH Terms
Adenosine Triphosphatases/metabolism Chromatography, Affinity/methods DNA Topoisomerases, Type II/isolation & purification,metabolism Escherichia coli/enzymology Genetic Complementation Test Kinetics Novobiocin Sepharose
Chemicals
Novobiocin Sepharose Adenosine Triphosphatases DNA Topoisomerases, Type II
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Staudenbauer W L
Orr E
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23 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/9.15.3589
Published
1981-08-11
Pages
3589-603
Language
English
Region
England
NLM ID
0411011
PMCID
PMC327377
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/9.15.3589 PMC Full Text View journal details
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