Linked Life Data

9200708

Source:http://linkedlifedata.com/resource/pubmed/id/9200708

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
24
pubmed:dateCreated
1997-7-10
pubmed:abstractText
Rapid-reaction methods have been used previously to identify intermediates in the reaction of the EcoRV restriction endonuclease on oligonucleotide substrates. In this study, the pathway on macromolecular DNA was elucidated by using the quench-flow method to analyze EcoRV reactions on a plasmid with one recognition site. Some reactions were carried out by first allowing the EcoRV enzyme to bind nonspecifically to the DNA and then initiating DNA cleavage by adding magnesium ions. The subsequent transfer of the enzyme from nonspecific to specific sites was extremely rapid, at a random walk rate of at least 5 x 10(5) base pairs per second. The two strands of the DNA at the EcoRV recognition site were then cleaved sequentially, at rates that were faster than the turnover number of the enzyme. The rates recorded for the cleavage steps were direct measurements of phosphodiester hydrolysis, while the turnover is limited by the dissociation of the product cleaved in both strands. Other reactions were initiated by adding EcoRV and MgCl2 to the DNA: these revealed not only the processes observed in reactions starting from DNA-bound enzyme but also the bimolecular association of the protein with the plasmid. The association rate was limited by diffusion but its rate constant, 1.2 x 10(8) M(-1) s(-1), was unusually small for the binding of a protein to DNA. The slowness of this diffusion-controlled process may be due to a rapid oscillation of the protein between closed and open conformations, with only the open form capable of binding DNA.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jun
pubmed:issn
0006-2960
pubmed:author
pubmed:issnType
Print
pubmed:day
17
pubmed:volume
36
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
7567-76
pubmed:dateRevised
2009-9-29
pubmed:meshHeading
pubmed:year
1997
pubmed:articleTitle
Rapid-reaction analysis of plasmid DNA cleavage by the EcoRV restriction endonuclease.
pubmed:affiliation
Department of Biochemistry, University of Bristol, U.K.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't