Aequorin variants with improved bioluminescence properties

E. Dikici, X. Qu, L. Rowe, L. Millner, C. Logue, Sapna K Deo, M. Ensor, Sylvia Daunert

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

The photoprotein aequorin has been widely used as a bioluminescent label in immunoassays, for the determination of calcium concentrations in vivo, and as a reporter in cellular imaging. It is composed of apoaequorin (189 amino acid residues), the imidazopyrazine chromophore coelenterazine and molecular oxygen. The emission characteristics of aequorin can be changed by rational design of the protein to introduce mutations in its structure, as well as by substituting different coelenterazine analogues to yield semi-synthetic aequorins. Variants of aequorin were created by mutating residues His16, Met19, Tyr82, Trp86, Trp108, Phe113 and Tyr132. Forty-two aequorin mutants were prepared and combined with 10 different coelenterazine analogues in a search for proteins with different emission wavelengths, altered decay kinetics and improved stability. This spectral tuning strategy resulted in semi-synthetic photoprotein mutants with significantly altered bioluminescent properties.

Original languageEnglish
Pages (from-to)243-248
Number of pages6
JournalProtein Engineering, Design and Selection
Volume22
Issue number4
DOIs
StatePublished - Apr 1 2009
Externally publishedYes

Fingerprint

Aequorin
Bioluminescence
Proteins
Luminescent Proteins
Molecular oxygen
Chromophores
Amino acids
Labels
Calcium
Tuning
Imaging techniques
Wavelength
Kinetics
Immunoassay
Oxygen
Amino Acids
Mutation
coelenterazine

Keywords

  • Aequorin
  • Bioluminescence
  • Coelenterazine analogues
  • Emission wavelength
  • Mutagenesis

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Bioengineering
  • Molecular Biology

Cite this

Aequorin variants with improved bioluminescence properties. / Dikici, E.; Qu, X.; Rowe, L.; Millner, L.; Logue, C.; Deo, Sapna K; Ensor, M.; Daunert, Sylvia.

In: Protein Engineering, Design and Selection, Vol. 22, No. 4, 01.04.2009, p. 243-248.

Research output: Contribution to journalArticle

Dikici, E. ; Qu, X. ; Rowe, L. ; Millner, L. ; Logue, C. ; Deo, Sapna K ; Ensor, M. ; Daunert, Sylvia. / Aequorin variants with improved bioluminescence properties. In: Protein Engineering, Design and Selection. 2009 ; Vol. 22, No. 4. pp. 243-248.
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