title: Fluorescence of tryptophan residues in firefly luciferases and enzyme--substrate complexes.
abstract: Quenching of tryptophan fluorescence of Luciola mingrelica (single tryptophan residue, Trp-419) and Photinus pyralis (two tryptophan residues, Trp-417 and Trp-426) luciferases with different quenchers (I-, Cs+, acrylamide) was studied. The conserved Trp-417(419) residue was shown to be not accessible to charged particles, and positively and negatively charged amino acid residues are located in close vicinity to it. We found previously unreported effective energy transfer from this tryptophan to luciferin during the quenching of the tryptophan fluorescence. The distance between the luciferin molecule and Trp-417(419) was calculated: 11-15 and 12-17 A for P. pyralis and L. mingrelica luciferases, respectively. The role of the conserved Trp residue in the catalysis is discussed. ATP and AMP are also quenchers of the tryptophan fluorescence of the luciferases. In this case, an allosteric mechanism of the interaction of Trp-417(419) with an excess of ATP (AMP) is proposed.
authors.affiliation: School of Chemistry, Lomonosov Moscow State University, Moscow, 119899, Russia.
authors.forename: E A
authors.initials: EA
authors.lastname: Chudinova
authors.forename: E I
authors.initials: EI
authors.lastname: Dementieva
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: D000249:Adenosine Monophosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000494:Allosteric Regulation; D000818:Animals; D002384:Catalysis; D001517:Coleoptera / Q000201:enzymology*; D005453:Fluorescence; D008156:Luciferases / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D013379:Substrate Specificity; D014364:Tryptophan / Q000378:metabolism*
chemical_list: D000249:Adenosine Monophosphate; D014364:Tryptophan; D000255:Adenosine Triphosphate; D008156:Luciferases