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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
4
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pubmed:dateCreated |
1998-6-4
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pubmed:abstractText |
The refolding of Clostridium symbiosum glutamate dehydrogenase (GDH) involves the formation of an inactive structured monomeric intermediate prior to its concentration-dependent association. The structured monomer obtained after removal of guanidinium chloride was stable and competent for reconstitution into active hexamers. Site-directed mutagenesis of C. symbiosum gdh gene was performed to replace the residues Arg-61 and Phe-187 which are involved in subunit-subunit interactions, as determined by three-dimensional structure analysis. Heterologous over-expression in Escherichia coli of the double mutant (R61E/F187D) led to the production of a soluble protein with a molecular mass consistent with the monomeric form of clostridial GDH. This protein is catalytically inactive but cross-reacts with an anti-wild-type GDH antibody preparation. The double mutant R61E/F187D does not assemble into hexamers. The physical properties and the stability toward guanidinium chloride and urea of R61E/F187D were studied and compared to those of the structured monomeric intermediate.
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/9568903-15299820,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9568903-1553382,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9568903-1587267,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9568903-5119250,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9568903-6696727,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9568903-7030122,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9568903-7126591,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9568903-7487949,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9568903-7663347,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9568903-8263929,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9568903-8428625,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9568903-8566542,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9568903-8591026,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9568903-8774713,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9568903-8917616
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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 |
0961-8368
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
7
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
966-74
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:9568903-Anilino Naphthalenesulfonates,
pubmed-meshheading:9568903-Binding Sites,
pubmed-meshheading:9568903-Circular Dichroism,
pubmed-meshheading:9568903-Clostridium,
pubmed-meshheading:9568903-Computer Simulation,
pubmed-meshheading:9568903-Escherichia coli,
pubmed-meshheading:9568903-Fluorescence,
pubmed-meshheading:9568903-Glutamate Dehydrogenase,
pubmed-meshheading:9568903-Guanidine,
pubmed-meshheading:9568903-Molecular Weight,
pubmed-meshheading:9568903-Mutagenesis, Site-Directed,
pubmed-meshheading:9568903-Protein Conformation,
pubmed-meshheading:9568903-Protein Denaturation,
pubmed-meshheading:9568903-Protein Engineering,
pubmed-meshheading:9568903-Protein Folding,
pubmed-meshheading:9568903-Protein Structure, Secondary,
pubmed-meshheading:9568903-Recombinant Proteins,
pubmed-meshheading:9568903-Ultracentrifugation,
pubmed-meshheading:9568903-Urea
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pubmed:year |
1998
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pubmed:articleTitle |
A monomeric mutant of Clostridium symbiosum glutamate dehydrogenase: comparison with a structured monomeric intermediate obtained during refolding.
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pubmed:affiliation |
Dipartimento di Scienze Biochimiche, A. Rossi Fanelli, Università La Sapienza, Roma, Italy.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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