Identification of luciferyl adenylate and luciferyl coenzyme a synthesized by firefly luciferase.
The firefly luciferase reaction intermediate luciferyl adenylate was detected by RP-HPLC analysis when the luciferase reaction was performed under a nitrogen atmosphere. Although this compound is always specified as an intermediate in the light-production reaction, this is the first report of its identification by HPLC in a luciferase assay medium. Under a low-oxygen atmosphere, luciferase can catalyze the synthesis of luciferyl coenzyme A from luciferin, ATP, and coenzyme A, but in air dehydroluciferyl coenzyme A was produced. The luciferase-catalyzed synthesis of these coenzyme A derivatives may be a consequence of the postulated recent evolutionary origin of firefly luciferases from an ancestral acyl-coenzyme A synthetase.
Fraga
Hugo
H
LAQUIPAI, Departamento de Química, Universidade do Porto, R. Campo Alegre 687, 4169-007 Porto, Portugal.
Esteves da Silva
Joaquim C G
JC
Fontes
Rui
R
D000255:Adenosine Triphosphate / Q000737:chemistry; D000818:Animals; D005075:Biological Evolution; D002851:Chromatography, High Pressure Liquid; D003065:Coenzyme A / Q000096:biosynthesis* / Q000235:genetics; D001517:Coleoptera / Q000201:enzymology*; D049454:Firefly Luciferin / Q000737:chemistry; D005612:Freeze Drying; D006863:Hydrogen-Ion Concentration; D008156:Luciferases / Q000378:metabolism*; D008163:Luminescent Measurements; D011755:Pyrophosphatases / Q000737:chemistry; D013237:Stereoisomerism
D049454:Firefly Luciferin; D000255:Adenosine Triphosphate; D008156:Luciferases; D011755:Pyrophosphatases; D003065:Coenzyme A