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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
1
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pubmed:dateCreated |
1981-5-13
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pubmed:abstractText |
Papain was succinylated in order to increase its solubility above pH 8 so that proton NMR spectroscopy could be used to study the ionization of His-159 at the active site of the enzyme. The pH dependence of NMR spectra of catalytically active succinyl-papain and the methylthio derivative of the active-site cysteinyl residue of succinyl-papain (succinyl-papain-S-SCH3) were determined between pH 6 and 10. The pH dependence of the C epsilon 1 H resonance of His-159 in catalytically active succinyl-papain indicates that His-159 has a pK of about 8.6 in the catalytically active form of the enzyme. The position of this resonance in succinyl-papain-S-SCH3 indicates that when the active-site cysteinyl residue is methylthiolated, His-159 is completely deprotonated between pH 6 and 10. This result is taken as evidence for an imidazolium--thiolate ion-pair interaction between His-159 and Cys-25 wherein neutralization of the charge on the thiolate anion by methylthiolation would be expected to cause a marked decrease in the pK of His-159. A possible catalytic role for the ion pair in the acylation step in papain-catalyzed reactions is proposed wherein attack of a substrate by the imidazolium--thiolate ion pair is accompanied by an increase in the acidity of the imidazolium group that facilitates expulsion of the leaving group of the substrate.
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pubmed:grant | |
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 |
Jan
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pubmed:issn |
0006-2960
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
6
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pubmed:volume |
20
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
48-51
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pubmed:dateRevised |
2007-11-14
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pubmed:meshHeading |
pubmed-meshheading:7470479-Binding Sites,
pubmed-meshheading:7470479-Cysteine,
pubmed-meshheading:7470479-Histidine,
pubmed-meshheading:7470479-Hydrogen-Ion Concentration,
pubmed-meshheading:7470479-Magnetic Resonance Spectroscopy,
pubmed-meshheading:7470479-Papain,
pubmed-meshheading:7470479-Protein Binding
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pubmed:year |
1981
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pubmed:articleTitle |
Effect of cysteine-25 on the ionization of histidine-159 in papain as determined by proton nuclear magnetic resonance spectroscopy. Evidence for a his-159--Cys-25 ion pair and its possible role in catalysis.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
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