Linked Life Data

2611219

Source:http://linkedlifedata.com/resource/pubmed/id/2611219

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
19
pubmed:dateCreated
1990-3-6
pubmed:abstractText
We have investigated in fluorescence stopped-flow and temperature-jump experiments the EcoRI-catalyzed cleavage of synthetic palindromic tridecadeoxynucleotides which contain the EcoRI site but differ in the flanking sequences. The overall reaction can be resolved in several reactions which were analyzed by a nonlinear least-squares fitting procedure on the experimental data. The result of this analysis is a minimal scheme that describes the overall reaction in terms of the rate constants of the individual reactions. According to this scheme EcoRI and the tridecadeoxynucleotide substrates associate in the presence of Mg2+ in a nearly diffusion-controlled process. This is followed by a reaction which is or includes the cleavage of the first phosphodiester bond. There is no indication for a time-resolved conformational transition prior to catalysis. After cleavage of the first strand, dissociation of the nicked double strand can occur, which then rearranges to the original palindromic double-stranded substrate and is bound again by the enzyme. Alternatively, the nicked double strand can be cleaved in the second strand. This reaction is followed by product release from the enzyme. The magnitude of the individual rate constants depends on the substrate used; the differences explain the preference of EcoRI for substrates that contain AT as compared to GC base pairs next to the recognition site.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Sep
pubmed:issn
0006-2960
pubmed:author
pubmed:issnType
Print
pubmed:day
19
pubmed:volume
28
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
7879-88
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed:year
1989
pubmed:articleTitle
Fluorescence stopped-flow kinetics of the cleavage of synthetic oligodeoxynucleotides by the EcoRI restriction endonuclease.
pubmed:affiliation
Zentrum Biochemie, Medizinische Hochschule Hannover, West Germany.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't