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. 1986 Nov;83(21):8107–8111. doi: 10.1073/pnas.83.21.8107

Site-specific mutagenesis of the calcium-binding photoprotein aequorin

Frederick I Tsuji *, Satoshi Inouye †,‡, Toshio Goto §, Yoshiyuki Sakaki †
PMCID: PMC386876  PMID: 16593774

Abstract

The luminescent protein aequorin from the jellyfish Aequoria victoria emits light by an intramolecular reaction in the presence of a trace amount of Ca2+. In order to understand the mechanism of the reaction, a study of structure-function relationships was undertaken with respect to modifying certain of its amino acid residues. This was done by carrying out oligonucleotide-directed site-specific mutagenesis of apoaequorin cDNA and expressing the mutagenized cDNA in Escherichia coli. Amino acid substitutions were made at the three Ca2+-binding sites, the three cysteines, and a histidine in one of the hydrophobic regions. Subsequent assay of the modified aequorin showed that the Ca2+-binding sites, the cysteines, and probably the histidine all play a role in the bioluminescence reaction of aequorin.

Keywords: bioluminescence, oxygenase, oligonucleotide-directed mutagenesis, protein modification

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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