Abstract
In the previous study, we have found that G65C and I125T double mutant of alpha chaperonin homo-oligomer from a hyperthermophilic archaeum, Thermococcus sp. strain KS-1, lacks ATP-dependent protein refolding activity despite showing ATPase activity and the ability to bind the denatured proteins. In this study, we have characterized several mutant Thermococcus chaperonin homo-oligomers with the amino acid substitutions of Gly-65 or Ile-125. The results showed that amino acid residue at 65th position should be a small amino acid such as glycine or alanine for the ATP-dependent refolding activity. The alpha chaperonin homo-oligomers with amino acid substitution of Gly-65 by amino acids whose side chains are larger than the methyl group did not have ATP-dependent protein refolding activity, but exhibited an increase of the binding affinity for unfolded proteins in the presence of ATP or AMP-PNP. (c)2001 Elsevier Science.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Adenosine Triphosphatases / chemistry
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Adenosine Triphosphatases / genetics
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Adenosine Triphosphatases / metabolism
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Adenosine Triphosphate / metabolism*
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Adenylyl Imidodiphosphate / metabolism
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Amino Acid Substitution
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Archaeal Proteins / chemistry*
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Archaeal Proteins / genetics
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Archaeal Proteins / metabolism*
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Base Sequence
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Chaperonins / chemistry*
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Chaperonins / genetics
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Chaperonins / metabolism*
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Escherichia coli / genetics
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Glycine / chemistry
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Green Fluorescent Proteins
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Kinetics
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Luminescent Proteins / chemistry
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Luminescent Proteins / genetics
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Luminescent Proteins / metabolism
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Mutagenesis, Site-Directed
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Plasmids / genetics
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Protein Folding
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Protein Structure, Quaternary
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Thermococcus / genetics
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Thermococcus / metabolism
Substances
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Archaeal Proteins
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Luminescent Proteins
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Recombinant Fusion Proteins
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Green Fluorescent Proteins
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Adenylyl Imidodiphosphate
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Adenosine Triphosphate
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Adenosine Triphosphatases
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Chaperonins
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Glycine