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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
28
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pubmed:dateCreated |
1992-8-25
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pubmed:abstractText |
Barnase is found to have a series of subsites for binding its substrates that confers large rate enhancements. Ribonucleotide substrates of the type Zp0Gp1Xp2Y have been synthesized, where p is phosphate, X, Y, and Z are nucleosides, and G is guanosine. G occupies the primary specificity site. The most important subsite is for p2, followed by that for Y. There appears to be no subsite for the Z or p0 positions. Occupation of the subsite for p2 gives rise to a 1000-fold increase in kcat/Km, composed of a 100-fold increase in kcat and a 10-fold decrease in Km. The Y subsite gives rise to further 20-fold increase in kcat/Km. Rates approaching diffusion control for kcat/Km are observed. kcat for the dinucleotide monophosphate GpU = 0.55 s-1, and Km = 240 microM; this compares with 53 s-1 and 20 microM for GpUp, and 3.3 x 10(3) s-1 and 17 microM for GpApA (the best substrate tested). Cleavage occurs at the 3'-phosphate of guanosine in all cases. There are differences in base specificity at the two subsites for X and Y downstream of the scissile bond. The binding energies of different substrates have been analyzed using thermodynamic cycles. These show that the contributions of the X and Y sites are nonadditive.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Jul
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pubmed:issn |
0006-2960
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
21
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pubmed:volume |
31
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
6390-5
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:1633151-Bacillus,
pubmed-meshheading:1633151-Binding Sites,
pubmed-meshheading:1633151-Kinetics,
pubmed-meshheading:1633151-Oligonucleotides,
pubmed-meshheading:1633151-Ribonucleases,
pubmed-meshheading:1633151-Structure-Activity Relationship,
pubmed-meshheading:1633151-Substrate Specificity,
pubmed-meshheading:1633151-Thermodynamics
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pubmed:year |
1992
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pubmed:articleTitle |
Barnase has subsites that give rise to large rate enhancements.
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pubmed:affiliation |
Department of Chemistry, University of Cambridge, England.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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