Fluorescent properties of firefly luciferases and their complexes with luciferin

被引：9

作者：

Dementieva, EI ^{[1
]}

Fedorchuk, EA ^{[1
]}

Brovko, LY ^{[1
]}

Savitskii, AP ^{[1
]}

Ugarova, NN ^{[1
]}

机构：

[1] Moscow MV Lomonosov State Univ, Sch Chem, Moscow 119899, Russia

来源：

BIOSCIENCE REPORTS | 2000年 / 20卷 / 01期

基金：

俄罗斯基础研究基金会;

关键词：

firefly luciferase; tryptophan fluorescence; luciferin; energy transfer;

D O I：

10.1023/A:1005579016387

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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 quencher;, 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 Angstrom for P. pyralis and 12-17 Angstrom for L. mingrelica luciferases. The role of the conserved Trp residue in the catalysis is discussed.

引用

页码：21 / 30

页数：10

共 12 条

[1]

BROVKO LY, 1996, BIOLUMINESCENCE CHEM, P206

[2]

BROVKO LY, 1997, J FLUORESC, V7, pS139

[3] Structural basis for the activation of phenylalanine in the non-ribosomal biosynthesis of gramicidin S [J].

Conti, E ;

Stachelhaus, T ;

Marahiel, MA ;

Brick, P .

EMBO JOURNAL, 1997, 16 (14) :4174-4183

[4] Crystal structure of firefly luciferase throws light on a superfamily of adenylate-forming enzymes [J].

Conti, E ;

Franks, NP ;

Brick, P .

STRUCTURE, 1996, 4 (03) :287-298

[5]

DEMENTEVA EI, 1986, BIOCHEMISTRY-MOSCOW+, V51, P111

[6]

Dementieva EI, 1996, BIOCHEMISTRY-MOSCOW+, V61, P115

[7] LUCIFERASE FROM THE EAST EUROPEAN FIREFLY LUCIOLA-MINGRELICA - CLONING AND NUCLEOTIDE-SEQUENCE OF THE CDNA, OVEREXPRESSION IN ESCHERICHIA-COLI AND PURIFICATION OF THE ENZYME [J].

DEVINE, JH ;

KUTUZOVA, GD ;

GREEN, VA ;

UGAROVA, NN ;

BALDWIN, TO .

BIOCHIMICA ET BIOPHYSICA ACTA, 1993, 1173 (02) :121-132

[8]

GANDELMAN OA, 1990, BIOCHEMISTRY-MOSCOW+, V55, P785

[9] INVESTIGATION OF THE INTERACTION BETWEEN FIREFLY LUCIFERASE AND OXYLUCIFERIN OR ITS ANALOGS BY STEADY-STATE AND SUBNANOSECOND TIME-RESOLVED FLUORESCENCE [J].

GANDELMAN, OA ;

BROVKO, LY ;

CHIKISHEV, AY ;

SHKURINOV, AP ;

UGAROVA, NN .

JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY B-BIOLOGY, 1994, 22 (03) :203-209

[10]

Lakowicz J. R., 2010, PRINCIPLES FLUORESCE

← 1 2 →