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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
1-2
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pubmed:dateCreated |
1993-4-13
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pubmed:abstractText |
The 51-62 loop of T4 phage lysozyme was altered by site-directed mutagenesis to obtain maximal homology with the typical EF-hand motif. A Ca(2+)-binding site was designed and created by replacing both Gly-51 and Asn-53 with aspartic acid. The mutant T4 lysozyme (G51D/N53D) was expressed in Escherichia coli. The activity of the G51D/N53D-mutant was about 60% of that of the wild-type protein. This mutant can bind Ca2+ ions specifically, while the effective dissociation constant was essentially greater than that of the EF-hand proteins. Stability of the G51D/N53D-mutant apo-form to urea- or temperature-induced denaturation was the same as that of the wild-type protein. In the presence of Ca2+ ions in solution the stability of the mutant T4 phage lysozyme was less than that of the wild-type protein. It is suggested that the binding of Ca2+ by the mutant is accompanied by the considerable conformational changes in the 'corrected' loop, which can lead to the Ca(2+)-induced destabilization of the protein.
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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 |
Mar
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pubmed:issn |
0006-3002
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
5
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pubmed:volume |
1162
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
84-8
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pubmed:dateRevised |
2000-12-18
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pubmed:meshHeading |
pubmed-meshheading:8448199-Amino Acid Sequence,
pubmed-meshheading:8448199-Bacteriophage T4,
pubmed-meshheading:8448199-Base Sequence,
pubmed-meshheading:8448199-Binding Sites,
pubmed-meshheading:8448199-Calcium,
pubmed-meshheading:8448199-Calcium-Binding Proteins,
pubmed-meshheading:8448199-Enzyme Stability,
pubmed-meshheading:8448199-Escherichia coli,
pubmed-meshheading:8448199-Molecular Sequence Data,
pubmed-meshheading:8448199-Muramidase,
pubmed-meshheading:8448199-Mutagenesis, Insertional,
pubmed-meshheading:8448199-Protein Conformation,
pubmed-meshheading:8448199-Protein Denaturation,
pubmed-meshheading:8448199-Tryptophan
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pubmed:year |
1993
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pubmed:articleTitle |
Introduction of Ca(2+)-binding amino-acid sequence into the T4 lysozyme.
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pubmed:affiliation |
Institute of Protein Research, Academy of Sciences of Russia, Pushchino, Moscow Region.
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pubmed:publicationType |
Journal Article
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