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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
5
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pubmed:dateCreated |
1995-10-10
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pubmed:abstractText |
Anomalous NMR behavior of the hydroxyl proton resonance for Ser 31 has been reported for histidine-containing protein (HPr) from two microorganisms: Escherichia coli and Staphylococcus aureus. The unusual slow exchange and chemical shift exhibited by the resonance led to the proposal that the hydroxyl group is involved in a strong hydrogen bond. To test this hypothesis and to characterize the importance of such an interaction, a mutant in which Ser 31 is replaced by an alanine was generated in HPr from Escherichia coli. The activity, stability, and structure of the mutant HPr were assessed using a reconstituted assay system, analysis of solvent denaturation curves, and NMR, respectively. Substitution of Ser 31 yields a fully functional protein that is only slightly less stable (delta delta G(folding) = 0.46 +/- 0.15 kcal mol-1) than the wild type. The NMR results confirm the identity of the hydrogen bond acceptor as Asp 69 and reveal that it exists as the gauche- conformer in wild-type HPr in solution but exhibits conformational averaging in the mutant protein. The side chain of Asp 69 interacts with two main-chain amide proteins in addition to its interaction with the side chain of Ser 31 in the wild-type protein. These results indicate that removal of the serine has led to the loss of all three hydrogen bond interactions involving Asp 69, suggesting a cooperative network of interactions. A complete analysis of the thermodynamics was performed in which differences in side-chain hydrophobicity and conformational entropy between the two proteins are accounted for.(ABSTRACT TRUNCATED AT 250 WORDS)
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-1483471,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-1549615,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-1668721,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-1680393,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-1711212,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-1751501,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-2155859,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-2372550,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-2539855,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-3233195,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-3542036,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-3691523,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-3773761,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-3881765,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-6096101,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-6307308,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-7712285,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-7803390,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-8158637,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-8182748,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-8226757,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-8365407,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-8421502,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7663349-8515453
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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 |
May
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pubmed:issn |
0961-8368
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
4
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
936-44
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:7663349-Bacterial Proteins,
pubmed-meshheading:7663349-Base Sequence,
pubmed-meshheading:7663349-Enzyme Stability,
pubmed-meshheading:7663349-Escherichia coli,
pubmed-meshheading:7663349-Hydrogen Bonding,
pubmed-meshheading:7663349-Magnetic Resonance Spectroscopy,
pubmed-meshheading:7663349-Models, Molecular,
pubmed-meshheading:7663349-Molecular Sequence Data,
pubmed-meshheading:7663349-Mutagenesis,
pubmed-meshheading:7663349-Phosphoenolpyruvate Sugar Phosphotransferase System,
pubmed-meshheading:7663349-Protein Conformation,
pubmed-meshheading:7663349-Protein Denaturation,
pubmed-meshheading:7663349-Protein Structure, Secondary,
pubmed-meshheading:7663349-Serine,
pubmed-meshheading:7663349-Staphylococcus aureus,
pubmed-meshheading:7663349-Thermodynamics,
pubmed-meshheading:7663349-Urea
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pubmed:year |
1995
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pubmed:articleTitle |
Investigation of a side-chain-side-chain hydrogen bond by mutagenesis, thermodynamics, and NMR spectroscopy.
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pubmed:affiliation |
Department of Biochemistry, University of Washington, Seattle 98195, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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