Nucleotide sequence of part of<i>Photobacterium leiognathi</i>lux region

Illarionov, Boris; Протопопова, М. В.; Karginov, Vladimir A.; Mertvetsov, N. P.; Gitelson, J. I.

doi:10.1093/nar/16.20.9855

Cited by 19 publications

(16 citation statements)

References 1 publication

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Bioluminescent bacteria belonging to the genus Photobacterium contain an additional gene located between luxB and luxE in the lux operon. This gene now known as luxF, was originally designated luxG (47), and independently as luxN (48). The luxF gene encodes a 24-kDa nonfluorescent flavoprotein whose function is unknown at present but binds two molecules of an unusual flavin mononucleotide adduct (45,46,49).…”

Section: ␣45mentioning

confidence: 99%

The 1.5-Å Resolution Crystal Structure of Bacterial Luciferase in Low Salt Conditions

Fisher

Thompson

Thoden

et al. 1996

Journal of Biological Chemistry

145

173

View full text Add to dashboard Cite

Bacterial luciferase is a flavin monooxygenase that catalyzes the oxidation of a long-chain aldehyde and releases energy in the form of visible light. A new crystal form of luciferase cloned from Vibrio harveyi has been grown under low-salt concentrations, which diffract xrays beyond 1.5-Å resolution. The x-ray structure of bacterial luciferase has been refined to a conventional Rfactor of 18.2% for all recorded synchrotron data between 30.0 and 1.50-Å resolution. Bacterial luciferase is an ␣-␤ heterodimer, and the individual subunits fold into a single domain (␤/␣) 8 barrel. The high resolution structure reveals a non-prolyl cis peptide bond that forms between Ala 74 and Ala 75 in the ␣ subunit near the putative active site. This cis peptide bond may have functional significance for creating a cavity at the active site. Bacterial luciferase employs reduced flavin as a substrate rather than a cofactor. The structure presented was determined in the absence of substrates. A comparison of the structural similarities between luciferase and a nonfluorescent flavoprotein, which is expressed in the lux operon of one genus of bioluminescent bacteria, suggests that the two proteins originated from a common ancestor. However, the flavin binding sites of the nonfluorescent protein are likely not representative of the flavin binding site on luciferase. The structure presented here will furnish a detailed molecular model for all bacterial luciferases.Living organisms that radiate light have been captivating people throughout the ages. Bioluminescent organisms such as fireflies, glowworms, mushrooms, fish, or bacteria represent a diverse range of species, which are widely dispersed in nature (1, 2). The enzymes that catalyze the bioluminescence reactions are named luciferases, and in most cases, their substrates are designated luciferins. These enzymes comprise a large evolutionarily diverse group, and the chemistry they catalyze is quite varied. Indeed, the only common factors of these enzymes is the requirement of O 2 , which was first established by Robert Boyle (3) more than 3 centuries ago. Today, it is known that all luciferase reactions are oxidative processes that convert a substrate to an electronically excited intermediate. Light emission occurs when the excited-state intermediate reverts back to the ground state resulting in the final product.Luminous bacteria are the most abundant and widely distributed of all bioluminescent organisms and are found in marine, freshwater, and terrestrial environments. Bacterial luciferase has been studied extensively and is the best understood of all luciferases. The luciferase of luminous bacteria is a flavin monooxygenase. Bacterial luciferase is an uncommon flavoprotein in that it employs reduced flavin as a substrate rather than a tightly bound cofactor. The enzyme catalyzes the reaction of FMNH 2 , O 2 , and a long-chain aliphatic aldehyde to yield FMN, the aliphatic carboxylic acid and blue-green light. All bacterial luciferases studied so far appear to be homologous, and all...

show abstract

Section: ␣45mentioning

confidence: 99%

The 1.5-Å Resolution Crystal Structure of Bacterial Luciferase in Low Salt Conditions

Fisher

Thompson

Thoden

et al. 1996

Journal of Biological Chemistry

145

173

View full text Add to dashboard Cite

show abstract

“…Thereafter, a new gene was found only in the lux operon of Photobacterium species. These genes were designated as luxF (Mancini, Boylan, Soly, Graham & Meighen, 1988) in P. phosphoreum and luxG (Illarionov, Protopopova, Karginov, Mertvetsov & Gitelson, 1988) or luxABN (Baldwin, Devine, Henckel, Lin & Shadel, 1989) in P. leiognathi, and the nucleotide sequences of both genes were determined (Soly, Mancini, Ferri, Boylan & Meighen, 1988;Illarionov et al, 1988;Baldwin et al, 1989). It was concluded that FP~,~) is identical to luxF protein; i.e., the luxF gene codes for FP390, by means of comparison between FP3,~o and luxF proteins by the molecular weight, the aminoacid composition of each entire protein, of all five CNBr fragments and the partial amino-acid sequences of three large CNBr fragments .…”

Section: Fmnh_~ + 02 + Rcho ~ Rcooh +Fmn + Ho + Hrmentioning

confidence: 99%

“…The DNA sequence of luxF gene and flanking non-coding region from two strains were identical with one exception; the 190th nucleotide from the beginning of the luxF gene was reported to be T in NCMB 844 whereas C was found in IFO 13896. However, T might be reported erroneously in NCMB 844 because the corresponding nucleotide in the luxF [designated as luxG by Illarionov et al (1988) or IuxABN by Baldwin et al (1989)] gene from P. leiognathi was reported to be C. Although the 64th amino-acid residue of FP39 o is deduced to be Ser in NCMB 844, this residue is deduced to be Pro in IFO 13896 as in P. leiognathi (Illarionov et al, 1988;Baldwin et al, 1989). A mutation of Pro to Set or its inverse seems to occur infrequently, as replacement of Pro residues for other amino-acid residues often causes a drastic structural change in the protein.…”

Section: Nucleotide Sequence Of Luxf Gene Of P Phosphoreum (Ifo 1389mentioning

confidence: 99%

See 1 more Smart Citation

Structure of flavoprotein FP390 from a luminescent bacterium Photobacterium phosphoreum refined at 2.7 Å resolution

Kita

Kasai²,

Miyata³

et al. 1996

Acta Crystallogr D Biol Crystallogr

View full text Add to dashboard Cite

The three-dimensional structure of a flavoprotein, FP~,~o, from a luminescent bacterium, Photobacterium phosphoreum, solved by the molecular-replacement method, was refined to an R factor of 24.0% for 17433 independent reflections, from 6.0 to 2.7~, resolution, collected by synchrotron radiation. The asymmetric unit of the crystal (space group P4322, a=b=76.8 and c --242 A) contains two monomer molecules related by a non-crystallographic twofold axis to form a dimer. There are two Q-flavin [flavin mononucleotide (FMN) with myristic acid] molecules in FP3,~o monomer. One of them is located at the interface of dimer which is bound to both monomer and the another is at the molecular surface. The electron density of myristic acids of Qflavins at the dimer interface in both monomer are weak and unclear, showing the possibility that the Q-flavins bound in this site are not a single species but a mixture of two components, 6-(3"-myristic acid)-FMN and 6-(4"-myristic acid)-FMN.

show abstract

“…In all luminous bacteria that have been studied, these five genes are closely linked and luxA and luxB are flanked by the genes for the enzymes of the fatty acid reductase complex. In both P. phosphoreum and P. leiognathi there is another gene between luxB and luxE (19,22). The function of the product of this gene is not known.…”

mentioning

confidence: 99%

Cloning, organization, and expression of the bioluminescence genes of Xenorhabdus luminescens

1990

View full text Add to dashboard Cite

The lux genes of Xenorhabdus luminescens, a symbiont of the nematode Heterorhabditis bacteriophora, were cloned and expressed in Escherichia coli. The expression of these genes in E. coli was qualitatively similar to their expression in X. luminescens. The organization of the genes is similar to that found in the marine luminous bacteria. Hybridization studies with the DNA that codes for the two subunits of luciferase revealed considerable homology among all of the strains of X. luminescens and with the DNA of other species of luminous bacteria, but none with the nonluminous Xenorhabdus species. Gross DNA alterations such as insertions, deletions, or inversions do not appear to be involved in the generation of dim variants known as secondary forms.Xenorhabdus luminescens is a luminous bacterium in the family Enterobacteriaceae (3,17,38 (1,6,7,18). The primary form is generally isolated from infective nematodes, but upon prolonged culture in various media secondary variants appear. In contrast to the primary form, the secondary variants are dim and lack detectable protease, lipase, antibiotic activity, protein crystals, and red pigment (6). The two forms also exhibit differences in colony morphology and staining properties (18). The secondary forms are deficient in providing optimum conditions for nematode reproduction (1, 7, 16).The physical and biochemical properties of the luminous system of X. luminescens are similar to those found in other bioluminescent bacteria (9,31,36 hyde to oxidized flavin and the corresponding long-chain fatty acid. A fatty acid reductase complex is required for the generation and recycling of fatty acid to aldehyde (34), and an NAD(P)H:flavin oxidoreductase supplies the reduced flavin (20). In rich media, the luminescence of X. luminescens increases dramatically during the late logarithmic or stationary phase of growth (36). The generation of secondary variants which are dim is another level of control of the luminous system of X. luminescens.The bioluminescence (lux) genes from Vibrio harveyi (8), Vibrio fischeri (12), Photobacterium leiognathi (11), and Photobacterium phosphoreum (22) have been cloned and expressed in Escherichia coli. Five structural genes are required for light emission: luxC, luxD, and luxE encode the fatty acid reductase complex, and luxA and luxB encode the alpha and beta subunits of bacterial luciferase (12,13,25). In all luminous bacteria that have been studied, these five genes are closely linked and luxA and luxB are flanked by the genes for the enzymes of the fatty acid reductase complex. In both P. phosphoreum and P. leiognathi there is another gene between luxB and luxE (19,22). The function of the product of this gene is not known.In this paper we report the cloning of the lux genes of X. luminescens and their expression when introduced into E. coli. We also present data showing the organization of the X. luminescens lux genes, the structure of the lux genes from the secondary form, and the homology of the luxA and luxB genes of X. luminescens Hm to DN...

show abstract

Nucleotide sequence of part ofPhotobacterium leiognathilux region

Cited by 19 publications

References 1 publication

The 1.5-Å Resolution Crystal Structure of Bacterial Luciferase in Low Salt Conditions

The 1.5-Å Resolution Crystal Structure of Bacterial Luciferase in Low Salt Conditions

Structure of flavoprotein FP390 from a luminescent bacterium Photobacterium phosphoreum refined at 2.7 Å resolution

Cloning, organization, and expression of the bioluminescence genes of Xenorhabdus luminescens

Contact Info

Product

Resources

About