rdf:type |
|
lifeskim:mentions |
|
pubmed:issue |
18
|
pubmed:dateCreated |
1989-10-24
|
pubmed:abstractText |
A Ca2+ binding site like an EF-hand motif was designed and created in human lysozyme by replacing both Gln-86 and Ala-92 with aspartic acids by site-directed mutagenesis. The mutant human lysozyme (D86/92-lysozyme) was expressed and secreted by yeast. One Ca2+ was found to bind one molecule of the purified protein with the binding constant 5.0 x 10(6) M-1. The enzymatic activity of holo-D86/92-lysozyme against glycol chitin at 40 degrees C was 2-fold higher than that of the native lysozyme. Maximal activity of the holo-D86/92-lysozyme was observed at 80 degrees C, where its relative activity normalized to the value at 40 degrees C was 6-fold and 17-fold higher than those of the native and apoenzymes, respectively. The activities of the native lysozyme and apo-D86/92-lysozyme were maximum at 65 degrees C-70 degrees C. Moreover, D86/92-lysozyme was more stable against protease digestion than the native lysozyme. These results indicate that the creation of the calcium binding site like an EF-hand motif in the human lysozyme enhances its structural stability.
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-1142432,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-13650640,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-271968,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-3059346,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-3072018,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-3109419,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-3118211,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-3200317,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-3214551,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-3288200,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-3299367,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-3666156,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-3783715,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-3785375,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-3974698,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-4382800,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-4530293,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-4570606,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-4700463,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-5229853,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-5532231,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-5532232,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-6225933,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-6387910,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-6709045,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-6774718,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-6866082,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-7207627,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-7260958,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-7277500,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-7334520,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2674939-7378374
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pubmed:language |
eng
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pubmed:journal |
|
pubmed:citationSubset |
IM
|
pubmed:chemical |
|
pubmed:status |
MEDLINE
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pubmed:month |
Sep
|
pubmed:issn |
0027-8424
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pubmed:author |
|
pubmed:issnType |
Print
|
pubmed:volume |
86
|
pubmed:owner |
NLM
|
pubmed:authorsComplete |
Y
|
pubmed:pagination |
6903-7
|
pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:2674939-Amino Acid Sequence,
pubmed-meshheading:2674939-Binding Sites,
pubmed-meshheading:2674939-Calcium,
pubmed-meshheading:2674939-DNA,
pubmed-meshheading:2674939-Enzyme Stability,
pubmed-meshheading:2674939-Humans,
pubmed-meshheading:2674939-Kinetics,
pubmed-meshheading:2674939-Molecular Sequence Data,
pubmed-meshheading:2674939-Muramidase,
pubmed-meshheading:2674939-Mutation,
pubmed-meshheading:2674939-Plasmids,
pubmed-meshheading:2674939-Protein Conformation,
pubmed-meshheading:2674939-Protein Denaturation,
pubmed-meshheading:2674939-Saccharomyces cerevisiae,
pubmed-meshheading:2674939-Sequence Homology, Nucleic Acid,
pubmed-meshheading:2674939-Thermodynamics
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pubmed:year |
1989
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pubmed:articleTitle |
Design and creation of a Ca2+ binding site in human lysozyme to enhance structural stability.
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pubmed:affiliation |
Protein Engineering Research Institute, Osaka, Japan.
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pubmed:publicationType |
Journal Article,
Comparative Study
|