Extraction of Renilla-type luciferin from the calcium-activated photoproteins aequorin, mnemiopsin, and berovin.
Photoproteins, which emit light in an oxygen-independent intramolecular reaction initiated by calcium ions, have been isolated from several bioluminescent organisms, including the hydrozoan jellyfish Aequorea and the ctenophore Mnemiopsis. The system of a related anthozoan coelenterate, the sea pansy Renilla reniformis, however, is oxygen dependent, requiring two organic components, luciferin and luciferase. Previously published indirect evidence indicates that photoproteins may contain a Renilla-type luciferin. We have now extracted in high yield a Renilla-type luciferin from three photoproteins, aequorin (45% yield), mnemiopsin (98% yield), and berovin (85% yield). Photoprotein luciferin, released from the holoprotein by mercaptoethanol treatment and separated from apo-photoprotein by gel filtration, no longer responds to calcium but now requires luciferase and O2 for light production. Photoprotein luciferin is identical to Renilla luciferin with respect to reaction kinetics and bioluminescence spectral distribution. In view of these results, the generally accepted hypothesis that the photoprotein chromophore is a protein-stabilized hydroperoxide of luciferin must be modified. We believe, instead, that the chromophore is free luciferin and that oxygen is bound as an oxygenated derivative of an amino-acid side chain of the protein. We propose the general term "coelenterate luciferin" to describe the light-producing chromophore from all bioluminescent coelenterates and ctenophores.
D000331:Aequorin / Q000032:analysis; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D003063:Cnidaria / Q000032:analysis* / Q000187:drug effects / Q000378:metabolism; D049454:Firefly Luciferin / Q000302:isolation & purification*; D007700:Kinetics; D008027:Light; D008156:Luciferases / Q000378:metabolism; D010777:Photochemistry; D011506:Proteins / Q000032:analysis* / Q000378:metabolism
D011506:Proteins; D000331:Aequorin; D049454:Firefly Luciferin; D008156:Luciferases; D002118:Calcium