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 language | English |
|---|---|
| Pages (from-to) | 243-248 |
| Number of pages | 6 |
| Journal | Protein Engineering, Design and Selection |
| Volume | 22 |
| Issue number | 4 |
| DOIs | |
| State | Published - Apr 1 2009 |
| Externally published | Yes |
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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 journal › Article
}
TY - JOUR
T1 - Aequorin variants with improved bioluminescence properties
AU - Dikici, E.
AU - Qu, X.
AU - Rowe, L.
AU - Millner, L.
AU - Logue, C.
AU - Deo, Sapna K
AU - Ensor, M.
AU - Daunert, Sylvia
PY - 2009/4/1
Y1 - 2009/4/1
N2 - 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.
AB - 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.
KW - Aequorin
KW - Bioluminescence
KW - Coelenterazine analogues
KW - Emission wavelength
KW - Mutagenesis
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UR - http://www.scopus.com/inward/citedby.url?scp=65349162391&partnerID=8YFLogxK
U2 - 10.1093/protein/gzn083
DO - 10.1093/protein/gzn083
M3 - Article
C2 - 19168563
AN - SCOPUS:65349162391
VL - 22
SP - 243
EP - 248
JO - Protein Engineering, Design and Selection
JF - Protein Engineering, Design and Selection
SN - 1741-0126
IS - 4
ER -