title: Crystal structure of coelenterazine-binding protein from Renilla muelleri at 1.7 A: why it is not a calcium-regulated photoprotein.
abstract: Bioluminescence in the sea pansy Renilla involves two distinct proteins, a Ca2+-triggered coelenterazine-binding protein (CBP), and Renilla luciferase. CBP contains one tightly bound coelenterazine molecule, which becomes available for reaction with luciferase and O2 only subsequent to Ca2+ binding. CBP belongs to the EF-hand superfamily of Ca2+-binding proteins and contains three "EF-hand" Ca2+-binding sites. The overall spatial structure of recombinant selenomethionine-labeled CBP determined at 1.7 A, is found to approximate the protein scaffold characteristic of the class of Ca2+-regulated photoproteins. Photoproteins however, catalyze molecular oxygen addition to coelenterazine producing a 2-hydroperoxycoelenterazine intermediate, which is stabilized within the binding cavity in the absence of Ca2+. Addition of Ca2+ triggers the bioluminescence reaction. However in CBP this first step of oxygen addition is not allowed. The different amino acid environments and hydrogen bond interactions within the binding cavity, are proposed to account for the different properties of the two classes of proteins.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA 30602, USA.
authors.forename: Galina A
authors.initials: GA
authors.lastname: Stepanyuk
authors.forename: Zhi-Jie
authors.initials: ZJ
authors.lastname: Liu
authors.forename: Svetlana S
authors.initials: SS
authors.lastname: Markova
authors.forename: Ludmila A
authors.initials: LA
authors.lastname: Frank
authors.forename: John
authors.initials: J
authors.lastname: Lee
authors.forename: Eugene S
authors.initials: ES
authors.lastname: Vysotski
authors.forename: Bi-Cheng
authors.initials: BC
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000737:chemistry / Q000378:metabolism; D018360:Crystallography, X-Ray; D007093:Imidazoles / Q000737:chemistry* / Q000378:metabolism*; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011719:Pyrazines / Q000737:chemistry* / Q000378:metabolism*; D048790:Renilla / Q000737:chemistry* / Q000235:genetics; D016415:Sequence Alignment
chemical_list: D007093:Imidazoles; D008164:Luminescent Proteins; D011719:Pyrazines; C017144:coelenterazine; D002118:Calcium