• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of pnasPNASInfo for AuthorsSubscriptionsAboutThis Article
Proc Natl Acad Sci U S A. May 1985; 82(10): 3154–3158.
PMCID: PMC397733

Cloning and sequence analysis of cDNA for the luminescent protein aequorin.

Abstract

The luminescent jellyfish Aequorea contains a photoprotein, aequorin, which emits light by an intramolecular reaction in the presence of a trace amount of Ca2+. A cDNA library of Aequorea was constructed and clones carrying the cDNA for the Ca2+-dependent photoprotein were isolated by the method of colony hybridization using synthetic oligonucleotide probes. The primary structure of the protein deduced from the nucleotide sequence showed that the protein is composed of 189 amino acid residues and has three E-F hand structures that are characteristic for Ca2+-binding sites. The sequence also suggested that the protein has hydrophobic regions at which the protein may interact with its functional chromophore, coelenterazine.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.0M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • SHIMOMURA O, JOHNSON FH, SAIGA Y. Extraction, purification and properties of aequorin, a bioluminescent protein from the luminous hydromedusan, Aequorea. J Cell Comp Physiol. 1962 Jun;59:223–239. [PubMed]
  • Shimomura O, Johnson FH, Saiga Y. Microdetermination of Calcium by Aequorin Luminescence. Science. 1963 Jun 21;140(3573):1339–1340. [PubMed]
  • Johnsn FH, Shimomura O. Preparation and use of aequorin for rapid microdetermination of Ca 2+ in biological systems. Nat New Biol. 1972 Jun 28;237(78):287–288. [PubMed]
  • Shimomura O, Shimomura A. EDTA-binding and acylation of the Ca2+-sensitive photoprotein aequorin. FEBS Lett. 1982 Feb 22;138(2):201–204. [PubMed]
  • Shimomura O, Shimomura A. Effect of calcium chelators on the Ca2+-dependent luminescence of aequorin. Biochem J. 1984 Aug 1;221(3):907–910. [PMC free article] [PubMed]
  • Shimomura O, Johnson FH. Structure of the light-emitting moiety of aequorin. Biochemistry. 1972 Apr 25;11(9):1602–1608. [PubMed]
  • Shimomura O, Johnson FH, Morise H. Mechanism of the luminescent intramolecular reaction of aequorin. Biochemistry. 1974 Jul 30;13(16):3278–3286. [PubMed]
  • Shimomura O, Johnson FH. Peroxidized coelenterazine, the active group in the photoprotein aequorin. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2611–2615. [PMC free article] [PubMed]
  • Shimomura O, Johnson FH. Regeneration of the photoprotein aequorin. Nature. 1975 Jul 17;256(5514):236–238. [PubMed]
  • Norgard MV, Tocci MJ, Monahan JJ. On the cloning of eukaryotic total poly(A)-RNA populations in Escherichia coli. J Biol Chem. 1980 Aug 25;255(16):7665–7672. [PubMed]
  • Miyata T, Iwanaga S, Sakata Y, Aoki N, Takamatsu J, Kamiya T. Plasminogens Tochigi II and Nagoya: two additional molecular defects with Ala-600----Thr replacement found in plasmin light chain variants. J Biochem. 1984 Aug;96(2):277–287. [PubMed]
  • Miyata T, Hiranaga M, Umezu M, Iwanaga S. Amino acid sequence of the coagulogen from Limulus polyphemus hemocytes. J Biol Chem. 1984 Jul 25;259(14):8924–8933. [PubMed]
  • Miyata T, Iwanaga S, Sakata Y, Aoki N. Plasminogen Tochigi: inactive plasmin resulting from replacement of alanine-600 by threonine in the active site. Proc Natl Acad Sci U S A. 1982 Oct;79(20):6132–6136. [PMC free article] [PubMed]
  • Miyata T, Usui K, Iwanaga S. The amino acid sequence of coagulogen isolated from southeast Asian horseshoe crab, Tachypleus gigas. J Biochem. 1984 Jun;95(6):1793–1801. [PubMed]
  • Hunkapiller MW, Hewick RM, Dreyer WJ, Hood LE. High-sensitivity sequencing with a gas-phase sequenator. Methods Enzymol. 1983;91:399–413. [PubMed]
  • Wallace RB, Johnson MJ, Hirose T, Miyake T, Kawashima EH, Itakura K. The use of synthetic oligonucleotides as hybridization probes. II. Hybridization of oligonucleotides of mixed sequence to rabbit beta-globin DNA. Nucleic Acids Res. 1981 Feb 25;9(4):879–894. [PMC free article] [PubMed]
  • Okayama H, Berg P. High-efficiency cloning of full-length cDNA. Mol Cell Biol. 1982 Feb;2(2):161–170. [PMC free article] [PubMed]
  • Birnboim HC, Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. [PMC free article] [PubMed]
  • Maxam AM, Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. [PubMed]
  • Blinks JR, Prendergast FG, Allen DG. Photoproteins as biological calcium indicators. Pharmacol Rev. 1976 Mar;28(1):1–93. [PubMed]
  • Proudfoot NJ, Brownlee GG. 3' non-coding region sequences in eukaryotic messenger RNA. Nature. 1976 Sep 16;263(5574):211–214. [PubMed]
  • Sherman L, Dafni N, Lieman-Hurwitz J, Groner Y. Nucleotide sequence and expression of human chromosome 21-encoded superoxide dismutase mRNA. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5465–5469. [PMC free article] [PubMed]
  • Watterson DM, Sharief F, Vanaman TC. The complete amino acid sequence of the Ca2+-dependent modulator protein (calmodulin) of bovine brain. J Biol Chem. 1980 Feb 10;255(3):962–975. [PubMed]
  • Kretsinger RH, Nockolds CE. Carp muscle calcium-binding protein. II. Structure determination and general description. J Biol Chem. 1973 May 10;248(9):3313–3326. [PubMed]
  • Collins JH, Potter JD, Horn MJ, Wilshire G, Jackman N. The amino acid sequence of rabbit skeletal muscle troponin C: gene replication and homology with calcium-binding proteins from carp and hake muscle. FEBS Lett. 1973 Nov 1;36(3):268–272. [PubMed]
  • Kyte J, Doolittle RF. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. [PubMed]
  • Shimomura O, Johnson FH. Properties of the bioluminescent protein aequorin. Biochemistry. 1969 Oct;8(10):3991–3997. [PubMed]
  • Kohama Y, Shimomura O, Johnson FH. Molecular weight of the photoprotein aequorin. Biochemistry. 1971 Oct 26;10(22):4149–4152. [PubMed]
  • Hastings JW, Mitchell G, Mattingly PH, Blinks JR, Van Leeuwen M. Response of aequorin bioluminescence to rapid changes in calcium concentration. Nature. 1969 Jun 14;222(5198):1047–1050. [PubMed]
  • Shimomura O, Johnson FH. Calcium binding, quantum yield, and emitting molecule in aequorin bioluminescence. Nature. 1970 Sep 26;227(5265):1356–1357. [PubMed]
  • Pless DD, Lennarz WJ. Enzymatic conversion of proteins to glycoproteins. Proc Natl Acad Sci U S A. 1977 Jan;74(1):134–138. [PMC free article] [PubMed]
  • Prasher D, McCann RO, Cormier MJ. Cloning and expression of the cDNA coding for aequorin, a bioluminescent calcium-binding protein. Biochem Biophys Res Commun. 1985 Feb 15;126(3):1259–1268. [PubMed]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...