| A new firefly luciferase with bimodal spectrum: identification of structural determinants of spectral pH-sensitivity in firefly luciferases. | |
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MedLine Citation:
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PMID: 16124832 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Fireflies 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. |
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Authors:
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Vadim R Viviani; T L Oehlmeyer; F G C Arnoldi; M R Brochetto-Braga |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Photochemistry and photobiology Volume: 81 ISSN: 0031-8655 ISO Abbreviation: Photochem. Photobiol. Publication Date: 2005 Jul-Aug |
Date Detail:
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Created Date: 2005-08-29 Completed Date: 2005-11-07 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 0376425 Medline TA: Photochem Photobiol Country: United States |
Other Details:
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Languages: eng Pagination: 843-8 Citation Subset: IM |
Affiliation:
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Departamento de Biologia, Instituto de Biociências, Universidade Estadual de São Paulo, Rio Claro, SP, Brazil. viviani@rc.unesp.br |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Sequence Animals Consensus Sequence Fireflies Hydrogen-Ion Concentration Kinetics Luciferases / chemistry*, metabolism Luminescent Measurements Luminescent Proteins Molecular Sequence Data Sequence Alignment |
| Chemical | |
Reg. No./Substance:
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0/Luminescent Proteins; EC 1.13.12.-/Luciferases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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