title: Fluorescent properties of firefly luciferases and their complexes with luciferin.
abstract: Fluorescence of luciferases from Luciola mingrelica (single tryptophan residue, Trp-419) and Photinus pyralis (two tryptophan residues, Trp-417, Trp-426) was studied. Analysis of quenching of tryptophan fluorescence showed that the tryptophan residue conserved in all luciferases is not accessible for charged quenchers, which is explained by the presence of positively and negatively charged amino acid residues in the close vicinity to it. An effective energy transfer from tryptophan to luciferin was observed during quenching of tryptophan fluorescence of both luciferases with luciferin. From the data on the energy transfer, the distance between the luciferin molecule and Trp-417 (419) in the luciferin luciferase complex was calculated: 11-15 A for P. pyralis and 12-17 A for L. mingrelica luciferases. The role of the conserved Trp residue in the catalysis is discussed.
authors.affiliation: M.V. Lomonosov Moscow State University, School of Chemistry, Russia. eid@enzyme.chem.msu.ru
authors.forename: E I
authors.initials: EI
authors.lastname: Dementieva
authors.forename: E A
authors.initials: EA
authors.lastname: Fedorchuk
authors.forename: L Y
authors.initials: LY
authors.lastname: Brovko
authors.forename: A P
authors.initials: AP
authors.lastname: Savitskii
authors.forename: N N
authors.initials: NN
authors.lastname: Ugarova
mesh_terms: D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D001517:Coleoptera / Q000201:enzymology; D049454:Firefly Luciferin / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D008156:Luciferases / Q000737:chemistry* / Q000378:metabolism; D008956:Models, Chemical; D008958:Models, Molecular; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D014364:Tryptophan / Q000737:chemistry / Q000378:metabolism
chemical_list: D000596:Amino Acids; D011994:Recombinant Proteins; D049454:Firefly Luciferin; D014364:Tryptophan; D008156:Luciferases