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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
18
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pubmed:dateCreated |
2004-4-26
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pubmed:databankReference |
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pubmed:abstractText |
Hint, histidine triad nucleotide-binding protein, is a universally conserved enzyme that hydrolyzes AMP linked to lysine and, in yeast, functions as a positive regulator of the RNA polymerase II C-terminal domain kinase, Kin28. To explore the biochemical and structural bases for the adenosine phosphoramidate hydrolase activity of rabbit Hint, we synthesized novel substrates linking a p-nitroaniline group to adenylate (AMP-pNA) and inhibitors that consist of an adenosine group and 5'-sulfamoyl (AdoOSO(2)NH(2)) or N-ethylsulfamoyl (AdoOSO(2)NHCH(2)CH(3)) group. AMP-pNA is a suitable substrate for Hint that allowed characterization of the inhibitors; titration of each inhibitor into AMP-pNA assays revealed their K(i) values. The N-ethylsulfamoyl derivative has a 13-fold binding advantage over the sulfamoyl adenosine. The 1.8-A cocrystal structure of rabbit Hint with N-ethylsulfamoyl adenosine revealed a binding site for the ethyl group against Trp-123, a residue that reaches across the Hint dimer interface to interact with the alkyl portion of the inhibitor and, presumably, the alkyl portion of a lysyl substrate. Ser-107 is positioned to donate a hydrogen bond to the leaving group nitrogen. Consistent with a role in acid-base catalysis, the Hint S107A mutant protein displayed depressed catalytic activity.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/14982931-10671479,
http://linkedlifedata.com/resource/pubmed/commentcorrection/14982931-10820011,
http://linkedlifedata.com/resource/pubmed/commentcorrection/14982931-10958787,
http://linkedlifedata.com/resource/pubmed/commentcorrection/14982931-11029061,
http://linkedlifedata.com/resource/pubmed/commentcorrection/14982931-11586299,
http://linkedlifedata.com/resource/pubmed/commentcorrection/14982931-11586300,
http://linkedlifedata.com/resource/pubmed/commentcorrection/14982931-11805111,
http://linkedlifedata.com/resource/pubmed/commentcorrection/14982931-12119013,
http://linkedlifedata.com/resource/pubmed/commentcorrection/14982931-12620103,
http://linkedlifedata.com/resource/pubmed/commentcorrection/14982931-12748294,
http://linkedlifedata.com/resource/pubmed/commentcorrection/14982931-12810953,
http://linkedlifedata.com/resource/pubmed/commentcorrection/14982931-1695712,
http://linkedlifedata.com/resource/pubmed/commentcorrection/14982931-5791926,
http://linkedlifedata.com/resource/pubmed/commentcorrection/14982931-9164465,
http://linkedlifedata.com/resource/pubmed/commentcorrection/14982931-9323207
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Apr
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pubmed:issn |
0021-9258
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
30
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pubmed:volume |
279
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
18711-6
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pubmed:dateRevised |
2010-12-3
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pubmed:meshHeading |
pubmed-meshheading:14982931-Adenosine Monophosphate,
pubmed-meshheading:14982931-Amino Acid Substitution,
pubmed-meshheading:14982931-Animals,
pubmed-meshheading:14982931-Binding Sites,
pubmed-meshheading:14982931-Carrier Proteins,
pubmed-meshheading:14982931-Crystallography, X-Ray,
pubmed-meshheading:14982931-Enzyme Inhibitors,
pubmed-meshheading:14982931-Hydrolases,
pubmed-meshheading:14982931-Kinetics,
pubmed-meshheading:14982931-Rabbits,
pubmed-meshheading:14982931-Substrate Specificity
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pubmed:year |
2004
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pubmed:articleTitle |
Biochemical, crystallographic, and mutagenic characterization of hint, the AMP-lysine hydrolase, with novel substrates and inhibitors.
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pubmed:affiliation |
Structural Biology and Bioinformatics Program, Kimmel Cancer Center, Philadelphia, Pennsylvania 19107, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, U.S. Gov't, Non-P.H.S.,
Research Support, Non-U.S. Gov't
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