Expression and characterization of the calcium-activated photoprotein from the ctenophore Bathocyroe fosteri: Insights into light-sensitive photoproteins

► We have expressed and characterized the first photoprotein from Bathocyroe fosteri. ► Emission spectra of BfosPP was at the upper limit of previous in vivo observations. ► This photoprotein is inactivated when exposed to certain wavelengths of light. ► There are non-conservative substitutions in t...

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Bibliographic Details
Published in:Biochemical and biophysical research communications Vol. 431; no. 2; pp. 360 - 366
Main Authors: Powers, Meghan L., McDermott, Amy G., Shaner, Nathan C., Haddock, Steven H.D.
Format: Journal Article
Language:English
Published: United States Elsevier Inc 08.02.2013
Subjects:
Amino Acid Sequence
Animals
Bioluminescence
blue light
Calcium - metabolism
Calcium-activated photoproteins
Calcium-Binding Proteins - chemistry
Calcium-Binding Proteins - classification
Calcium-Binding Proteins - genetics
Calcium-Binding Proteins - radiation effects
Cloning, Molecular
color
Ctenophora
Ctenophora - metabolism
Ctenophore
Imidazoles - chemistry
Luminescence
Luminescent Agents - chemistry
luminescent proteins
Luminescent Proteins - chemistry
Luminescent Proteins - classification
Luminescent Proteins - genetics
Luminescent Proteins - radiation effects
Molecular Sequence Data
Phylogeny
prediction
Protein Conformation
Pyrazines - chemistry
sequence homology
wavelengths
Ctenophore
Bioluminescence
Calcium-activated photoproteins
ISSN:0006-291X, 1090-2104, 1090-2104
Online Access:Get full text
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Summary:► We have expressed and characterized the first photoprotein from Bathocyroe fosteri. ► Emission spectra of BfosPP was at the upper limit of previous in vivo observations. ► This photoprotein is inactivated when exposed to certain wavelengths of light. ► There are non-conservative substitutions in the putative binding-pocket of ctenophores. Calcium-binding photoproteins have been discovered in a variety of luminous marine organisms [1]. Recent interest in photoproteins from the phylum Ctenophora has stemmed from cloning and expression of several photoproteins from this group [2–5]. Additional characterization has revealed unique biochemical properties found only in ctenophore photoproteins, such as inactivation by light. Here we report the cloning, expression, and characterization of the photoprotein responsible for luminescence in the deep-sea ctenophore Bathocyroefosteri. This animal was of particular interest due to the unique broad color spectrum observed in live specimens [6]. Full-length sequences were identified by BLAST searches of known photoprotein sequences against Bathocyroe transcripts obtained from 454 sequencing. Recombinantly expressed Bathocyroe photoprotein (BfosPP) displayed an optimal coelenterazine-loading pH of 8.5, and produced calcium-triggered luminescence with peak wavelengths closely matching the 493nm peak observed in the spectrum of live B.fosteri specimens. Luminescence from recombinant BfosPP was inactivated most efficiently by UV and blue light. Primary structure alignment of BfosPP with other characterized photoproteins showed very strong sequence similarity to other ctenophore photoproteins and conservation of EF-hand motifs. Both alignment and structural prediction data provide more insight into the formation of the coelenterazine-binding domain and the probable mechanism of photoinactivation.
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ISSN:0006-291X
1090-2104
1090-2104
DOI:10.1016/j.bbrc.2012.12.026