Active-site properties of Phrixotrix railroad worm green and red bioluminescence-eliciting luciferases
Contribuinte(s) |
Universidade Estadual Paulista (UNESP) |
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Data(s) |
20/05/2014
20/05/2014
01/10/2006
|
Resumo |
The luciferases of the railroad worm Phrixotrix (Coleoptera: Phengodidae) are the only beetle luciferases that naturally produce true red bioluminescence. Previously, we cloned the green- (PxGR) and red-emitting (PxRE) luciferases of railroad worms Phrixotrix viviani and P. hirtus[OLE1]. These luciferases were expressed and purified, and their active-site properties were determined. The red-emitting PxRE luciferase displays flash-like kinetics, whereas PxGR luciferase displays slow-type kinetics. The substrate affinities and catalytic efficiency of PxRE luciferase are also higher than those of PxGR luciferase. Fluorescence studies with 8-anilino-1-naphthalene sulfonic acid and 6-p-toluidino-2-naphthalene sulfonic acid showed that the PxRE luciferase luciferin-binding site is more polar than that of PxGR luciferase, and it is sensitive to guanidine. Alutagenesis and modelling studies suggest that several invariant residues in the putative luciferin-binding site of PxRE luciferase cannot interact with excited oxyluciferin. These results suggest that one portion of the luciferin-binding site of the red-emitting luciferase is tighter than that of PxGR luciferase, whereas the other portion could be more open and polar. |
Formato |
467-474 |
Identificador |
http://dx.doi.org/10.1093/jb/mvj190 Journal of Biochemistry. Tokyo: Japanese Biochemical Soc, v. 140, n. 4, p. 467-474, 2006. 0021-924X http://hdl.handle.net/11449/19562 10.1093/jb/mvj190 WOS:000241798200001 WOS000241798200001.pdf |
Idioma(s) |
eng |
Publicador |
Japanese Biochemical Soc |
Relação |
Journal of Biochemistry |
Direitos |
closedAccess |
Palavras-Chave | #bioluminescence #luciferases #Phrixotrix #railroad worms |
Tipo |
info:eu-repo/semantics/article |