title: Expression and characterization of the calcium-activated photoprotein from the ctenophore Bathocyroe fosteri: insights into light-sensitive photoproteins.
abstract: 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 Bathocyroe fosteri. 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 493 nm 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.
authors.affiliation: Monterey Bay Aquarium Research Institute, 7700 Sandholdt Road, Moss Landing, CA 95039, USA. mpowers@mbari.org
authors.forename: Meghan L
authors.initials: ML
authors.lastname: Powers
authors.forename: Amy G
authors.initials: AG
authors.lastname: McDermott
authors.forename: Nathan C
authors.initials: NC
authors.lastname: Shaner
authors.forename: Steven H D
authors.initials: SH
authors.lastname: Haddock
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000737:chemistry* / Q000145:classification / Q000235:genetics / Q000528:radiation effects*; D003001:Cloning, Molecular; D037302:Ctenophora / Q000378:metabolism*; D007093:Imidazoles / Q000737:chemistry; D049449:Luminescence; D049408:Luminescent Agents / Q000737:chemistry; D008164:Luminescent Proteins / Q000737:chemistry* / Q000145:classification / Q000235:genetics / Q000528:radiation effects*; D008969:Molecular Sequence Data; D010802:Phylogeny; D011487:Protein Conformation; D011719:Pyrazines / Q000737:chemistry
chemical_list: D002135:Calcium-Binding Proteins; D007093:Imidazoles; D049408:Luminescent Agents; D008164:Luminescent Proteins; D011719:Pyrazines; C017144:coelenterazine; D002118:Calcium