A new firefly luciferase with bimodal spectrum: Identification of structural determinants of spectral pH-sensitivity in firefly luciferases

dc.contributor.authorViviani, Vadim R. [UNESP]
dc.contributor.authorOehlmeyer, T. L. [UNESP]
dc.contributor.authorArnoldi, F. G C [UNESP]
dc.contributor.authorBrochetto-Braga, M. R. [UNESP]
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.date.accessioned2014-05-27T11:21:22Z
dc.date.available2014-05-27T11:21:22Z
dc.date.issued2005-07-01
dc.description.abstractFireflies emit flashes in the green-yellow region of the spectrum for the purpose of sexual attraction. The bioluminescence color is determined by the luciferases. It is well known that the in vitro bioluminescence color of firefly luciferases can be shifted toward the red by lower pH and higher temperature; for this reason they are classified as pH-sensitive luciferases. However, the mechanism and structural origin of pH sensitivity in fireflies remains unknown. Here we report the cloning of a new luciferase from the Brazilian twilight active firefly Macrolampis sp2, which displays an unusual bimodal spectrum. The recombinant luciferase displays a sensitive spectrum with the peak at 569 nm and a shoulder in the red region. Comparison of the bioluminescence spectra of Macrolampis, Photinus and Cratomorphus firefly luciferases shows that the distinct colors are determined by the ratio between green and red emitters under luciferase influence. Comparison of Macrolampis luciferase with the highly similar North American Photinus pyralis luciferase (91%) showed few substitutions potentially involved with the higher spectral sensitivity in Macrolampis luciferase. Site-directed mutagenesis showed that the natural substitution E354N determines the appearance of the shoulder in the red region of Macrolampis luciferase bioluminescence spectrum, helping to identify important interactions and residues involved in the pH-sensing mechanism in firefly luciferases. © 2005 American Society for Photobiology.en
dc.description.affiliationDepartamento de Biologia Instituto de Biociências Universidade Estadual de São Paulo (UNESP), Rio Claro, SP
dc.description.affiliationDepartamento de Biologia Instituto de Biociências Universidade Estadual de São Paulo, Rio Claro, SP, 13506-900
dc.description.affiliationUnespDepartamento de Biologia Instituto de Biociências Universidade Estadual de São Paulo (UNESP), Rio Claro, SP
dc.description.affiliationUnespDepartamento de Biologia Instituto de Biociências Universidade Estadual de São Paulo, Rio Claro, SP, 13506-900
dc.format.extent843-848
dc.identifierhttp://dx.doi.org/10.1562/2004-12-09-RA-398R.1
dc.identifier.citationPhotochemistry and Photobiology, v. 81, n. 4, p. 843-848, 2005.
dc.identifier.doi10.1562/2004-12-09-RA-398R.1
dc.identifier.issn0031-8655
dc.identifier.scopus2-s2.0-24644484204
dc.identifier.urihttp://hdl.handle.net/11449/68297
dc.language.isoeng
dc.relation.ispartofPhotochemistry and Photobiology
dc.relation.ispartofjcr2.214
dc.relation.ispartofsjr0,591
dc.rights.accessRightsAcesso restrito
dc.sourceScopus
dc.subjectluciferase
dc.subjectphotoprotein
dc.subjectamino acid sequence
dc.subjectanimal
dc.subjectchemistry
dc.subjectconsensus sequence
dc.subjectfirefly
dc.subjectkinetics
dc.subjectluminescence
dc.subjectmetabolism
dc.subjectmolecular genetics
dc.subjectpH
dc.subjectsequence alignment
dc.subjectAmino Acid Sequence
dc.subjectAnimals
dc.subjectConsensus Sequence
dc.subjectFireflies
dc.subjectHydrogen-Ion Concentration
dc.subjectKinetics
dc.subjectLuciferases
dc.subjectLuminescent Measurements
dc.subjectLuminescent Proteins
dc.subjectMolecular Sequence Data
dc.subjectSequence Alignment
dc.subjectLampyridae
dc.subjectPhotinus
dc.subjectPhotinus pyralis
dc.titleA new firefly luciferase with bimodal spectrum: Identification of structural determinants of spectral pH-sensitivity in firefly luciferasesen
dc.typeArtigo
dcterms.licensehttp://www.bioone.org/page/terms_of_use
unesp.campusUniversidade Estadual Paulista (Unesp), Instituto de Biociências, Rio Claropt

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