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rdf:type |
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lifeskim:mentions |
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pubmed:dateCreated |
1993-8-12
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pubmed:abstractText |
cDNA coding for the Ca(2+)-activated photoprotein aequorin from the jellyfish Aequorea victoria has been engineered to investigate the role of the C-terminal proline residue in bioluminescence. Recombinant aequorin proteins were synthesized by PCR followed by in vitro transcription/translation, and characterized by specific activity, stability, and affinity for coelenterazine. The C-terminal proline residue of aequorin was shown to be essential for the long-term stability of the bound coelenterazine. Aequorin minus proline had only 1% of the specific activity of the wild-type after 2 h, and was virtually inactive after 18 h. The instability of this variant was further demonstrated by re-activating with a coelenterazine analogue (epsilon-coelenterazine), where maximum reactivation was reached in 15 min, and the luminescent activity was almost completely abolished within 3 h. Replacement of the C-terminal proline residue with histidine or glutamic acid decreased the specific activity to 10 and 19% of that of the wild-type respectively. However these variants were also unstable, having t1/2 values of 2.4 h and 2.3 h respectively. Enhancement of the Ca(2+)-independent light emission when proline was replaced by histidine confirmed the stabilizing role of the C-terminal proline. No significant effect of removal of the C-terminal proline was detected on the affinity for coelenterazine.
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/8101077-14203166,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8101077-1530606,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8101077-16593774,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8101077-1765170,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8101077-1953665,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8101077-1970919,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8101077-2241947,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8101077-2479149,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8101077-275832,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8101077-2801233,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8101077-2866797,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8101077-3340835,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8101077-3401214,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8101077-3858813,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8101077-5432063,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8101077-841325
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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 |
http://linkedlifedata.com/resource/pubmed/chemical/Aequorin,
http://linkedlifedata.com/resource/pubmed/chemical/Calcium,
http://linkedlifedata.com/resource/pubmed/chemical/DNA,
http://linkedlifedata.com/resource/pubmed/chemical/Glutamates,
http://linkedlifedata.com/resource/pubmed/chemical/Glutamic Acid,
http://linkedlifedata.com/resource/pubmed/chemical/Histidine,
http://linkedlifedata.com/resource/pubmed/chemical/Imidazoles,
http://linkedlifedata.com/resource/pubmed/chemical/Proline,
http://linkedlifedata.com/resource/pubmed/chemical/Pyrazines,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/coelenterazine
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pubmed:status |
MEDLINE
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pubmed:month |
Jul
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pubmed:issn |
0264-6021
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
1
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pubmed:volume |
293 ( Pt 1)
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
181-5
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pubmed:dateRevised |
2010-9-10
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pubmed:meshHeading |
pubmed-meshheading:8101077-Aequorin,
pubmed-meshheading:8101077-Animals,
pubmed-meshheading:8101077-Base Sequence,
pubmed-meshheading:8101077-Calcium,
pubmed-meshheading:8101077-DNA,
pubmed-meshheading:8101077-Electrophoresis, Agar Gel,
pubmed-meshheading:8101077-Glutamates,
pubmed-meshheading:8101077-Glutamic Acid,
pubmed-meshheading:8101077-Histidine,
pubmed-meshheading:8101077-Imidazoles,
pubmed-meshheading:8101077-Luminescent Measurements,
pubmed-meshheading:8101077-Molecular Sequence Data,
pubmed-meshheading:8101077-Mutation,
pubmed-meshheading:8101077-Polymerase Chain Reaction,
pubmed-meshheading:8101077-Proline,
pubmed-meshheading:8101077-Protein Biosynthesis,
pubmed-meshheading:8101077-Pyrazines,
pubmed-meshheading:8101077-Recombinant Proteins,
pubmed-meshheading:8101077-Scyphozoa
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pubmed:year |
1993
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pubmed:articleTitle |
Requirement of the C-terminal proline residue for stability of the Ca(2+)-activated photoprotein aequorin.
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pubmed:affiliation |
Department of Medical Biochemistry, University of Wales College of Medicine, Heath Park, Cardiff, U.K.
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pubmed:publicationType |
Journal Article,
Comparative Study,
In Vitro,
Research Support, Non-U.S. Gov't
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