title: Interchange of aequorin and obelin bioluminescence color is determined by substitution of one active site residue of each photoprotein.
abstract: The bioluminescence spectra from the Ca2+-regulated photoproteins aequorin (lambdamax=469 nm) and obelin (lambdamax=482 nm) differ because aequorin has an H-bond from its Tyr82 to the bound coelenteramide, not present in obelin at the corresponding Phe88. Substitutions of this Phe88 by Tyr, Trp, or His shifted the obelin bioluminescence to shorter wavelength with F88Y having lambdamax=453 nm. Removal of the H-bond by the substitution of Y82F in aequorin shifted its bioluminescence to lambdamax=501 nm. All mutants were stable with good activity and were expressible in mammalian cells, thereby demonstrating potential for monitoring multiple events in cells using multi-color detection.
authors.affiliation: Photobiology Laboratory, Institute of Biophysics, Siberian Branch, Russian Academy of Sciences, Krasnoyarsk 660036, Russia.
authors.forename: Galina A
authors.initials: GA
authors.lastname: Stepanyuk
authors.forename: Stefan
authors.initials: S
authors.lastname: Golz
authors.forename: Svetlana V
authors.initials: SV
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
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology; D000331:Aequorin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D016466:CHO Cells; D002118:Calcium / Q000494:pharmacology; D003116:Color; D006224:Cricetinae; D008163:Luminescent Measurements; D008164:Luminescent Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D018048:Receptors, Purinergic P2 / Q000378:metabolism; D058473:Receptors, Purinergic P2Y2; D016415:Sequence Alignment; D013050:Spectrometry, Fluorescence
chemical_list: D008164:Luminescent Proteins; D018048:Receptors, Purinergic P2; D058473:Receptors, Purinergic P2Y2; C010497:obelin; D000331:Aequorin; D000255:Adenosine Triphosphate; D002118:Calcium