Matches in SemOpenAlex for { <https://semopenalex.org/work/W2047034905> ?p ?o ?g. }
- W2047034905 endingPage "8503" @default.
- W2047034905 startingPage "8499" @default.
- W2047034905 abstract "The crystal structure analyses of the EcoRI-DNA and EcoRV-DNA complexes do not provide clear suggestions as to which amino acid residues are responsible for the activation of water to carry out the DNA cleavage. Based on molecular modeling, we have proposed recently that the attacking water molecule is activated by the negatively charged pro-Rp phosphoryl oxygen of the phosphate group 3' to the scissile phosphodiester bond. We now present experimental evidence to support this proposal. (i) Oligodeoxynucleotide substrates lacking this phosphate group in one strand are cleaved only in the other strand. (ii) Oligodeoxynucleotide substrates carrying an H-phosphonate substitution at this position in both strands and, therefore, lacking a negatively charged oxygen at this position are cleaved at least four orders of magnitude more slowly than the unmodified substrate. These results are supported by other modification studies: oligodeoxynucleotide substrates with a phosphorothioate substitution at this position in both strands are cleaved only if the negatively charged sulfur is in the RP configuration as shown for EcoRI [Koziolkiewicz, M. & Stec, W.J. (1992) Biochemistry 31, 9460-9466] and EcoRV (B. A. Connolly, personal communication). As the phosphate residue 3' to the scissile phosphodiester bond is not needed for strong DNA binding by both enzymes, these findings strongly suggest that this phosphate group plays an active role during catalysis. This proposal, furthermore, gives a straightforward explanation of why in the EcoRI-DNA and EcoRV-DNA complexes the DNA is distorted differently, but in each case the 3' phosphate group closely approaches the phosphate group that is attacked. Finally, an alternative mechanism for DNA cleavage involving two metal ions is unlikely in the light of our finding that both EcoRI and EcoRV need only one Mg2+ per active site for cleavage." @default.
- W2047034905 created "2016-06-24" @default.
- W2047034905 creator A5033630820 @default.
- W2047034905 creator A5046166838 @default.
- W2047034905 creator A5055189220 @default.
- W2047034905 creator A5070638287 @default.
- W2047034905 creator A5088616795 @default.
- W2047034905 date "1993-09-15" @default.
- W2047034905 modified "2023-10-14" @default.
- W2047034905 title "Substrate-assisted catalysis in the cleavage of DNA by the EcoRI and EcoRV restriction enzymes." @default.
- W2047034905 cites W136740849 @default.
- W2047034905 cites W1485873149 @default.
- W2047034905 cites W1510656481 @default.
- W2047034905 cites W1519899019 @default.
- W2047034905 cites W1593479774 @default.
- W2047034905 cites W1594420006 @default.
- W2047034905 cites W1965753868 @default.
- W2047034905 cites W1980254677 @default.
- W2047034905 cites W1981929180 @default.
- W2047034905 cites W1982878977 @default.
- W2047034905 cites W1984541502 @default.
- W2047034905 cites W1998861841 @default.
- W2047034905 cites W2005398496 @default.
- W2047034905 cites W2006869216 @default.
- W2047034905 cites W2013998090 @default.
- W2047034905 cites W2023072985 @default.
- W2047034905 cites W2029344548 @default.
- W2047034905 cites W2030551452 @default.
- W2047034905 cites W2037088676 @default.
- W2047034905 cites W2045606892 @default.
- W2047034905 cites W2054312963 @default.
- W2047034905 cites W2061148784 @default.
- W2047034905 cites W2067448597 @default.
- W2047034905 cites W2068215202 @default.
- W2047034905 cites W2082391675 @default.
- W2047034905 cites W2091745929 @default.
- W2047034905 cites W2092171870 @default.
- W2047034905 cites W2093217670 @default.
- W2047034905 cites W2100067555 @default.
- W2047034905 cites W2102428701 @default.
- W2047034905 cites W2133174507 @default.
- W2047034905 cites W2211117662 @default.
- W2047034905 cites W4254085062 @default.
- W2047034905 doi "https://doi.org/10.1073/pnas.90.18.8499" @default.
- W2047034905 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/47384" @default.
- W2047034905 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/8378323" @default.
- W2047034905 hasPublicationYear "1993" @default.
- W2047034905 type Work @default.
- W2047034905 sameAs 2047034905 @default.
- W2047034905 citedByCount "110" @default.
- W2047034905 countsByYear W20470349052012 @default.
- W2047034905 countsByYear W20470349052013 @default.
- W2047034905 countsByYear W20470349052014 @default.
- W2047034905 countsByYear W20470349052018 @default.
- W2047034905 countsByYear W20470349052019 @default.
- W2047034905 countsByYear W20470349052020 @default.
- W2047034905 countsByYear W20470349052022 @default.
- W2047034905 crossrefType "journal-article" @default.
- W2047034905 hasAuthorship W2047034905A5033630820 @default.
- W2047034905 hasAuthorship W2047034905A5046166838 @default.
- W2047034905 hasAuthorship W2047034905A5055189220 @default.
- W2047034905 hasAuthorship W2047034905A5070638287 @default.
- W2047034905 hasAuthorship W2047034905A5088616795 @default.
- W2047034905 hasBestOaLocation W20470349052 @default.
- W2047034905 hasConcept C104317684 @default.
- W2047034905 hasConcept C128319531 @default.
- W2047034905 hasConcept C130123407 @default.
- W2047034905 hasConcept C181199279 @default.
- W2047034905 hasConcept C185592680 @default.
- W2047034905 hasConcept C2776777567 @default.
- W2047034905 hasConcept C38441620 @default.
- W2047034905 hasConcept C41183919 @default.
- W2047034905 hasConcept C552990157 @default.
- W2047034905 hasConcept C55493867 @default.
- W2047034905 hasConcept C67705224 @default.
- W2047034905 hasConcept C71240020 @default.
- W2047034905 hasConcept C87282627 @default.
- W2047034905 hasConceptScore W2047034905C104317684 @default.
- W2047034905 hasConceptScore W2047034905C128319531 @default.
- W2047034905 hasConceptScore W2047034905C130123407 @default.
- W2047034905 hasConceptScore W2047034905C181199279 @default.
- W2047034905 hasConceptScore W2047034905C185592680 @default.
- W2047034905 hasConceptScore W2047034905C2776777567 @default.
- W2047034905 hasConceptScore W2047034905C38441620 @default.
- W2047034905 hasConceptScore W2047034905C41183919 @default.
- W2047034905 hasConceptScore W2047034905C552990157 @default.
- W2047034905 hasConceptScore W2047034905C55493867 @default.
- W2047034905 hasConceptScore W2047034905C67705224 @default.
- W2047034905 hasConceptScore W2047034905C71240020 @default.
- W2047034905 hasConceptScore W2047034905C87282627 @default.
- W2047034905 hasIssue "18" @default.
- W2047034905 hasLocation W20470349051 @default.
- W2047034905 hasLocation W20470349052 @default.
- W2047034905 hasLocation W20470349053 @default.
- W2047034905 hasLocation W20470349054 @default.
- W2047034905 hasOpenAccess W2047034905 @default.
- W2047034905 hasPrimaryLocation W20470349051 @default.
- W2047034905 hasRelatedWork W1965500916 @default.