Bioluminescence of the arm light organs of the luminous squid Watasenia scintillans

• Published in: Biochimica et biophysica acta Vol. 1780; no. 5; pp. 784 - 792
• Main Authors: Teranishi, Katsunori, Shimomura, Osamu
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.05.2008
• Subjects: Adenosine Diphosphate - chemistry; Adenosine Triphosphate - analogs & derivatives; Adenosine Triphosphate - chemistry; Animal Structures - chemistry; Animal Structures - enzymology; Animals; ATP; Bioluminescence; Catalysis; Coelenterazine disulfate; Decapodiformes - chemistry; Decapodiformes - enzymology; Enzyme Stability; Hydrogen-Ion Concentration; Imidazoles - chemistry; Kinetics; Luciferase; Luciferases - chemistry; Luciferases - isolation & purification; Luciferases - metabolism; Luminescence; Luminescent Measurements; Magnesium Chloride - chemistry; Molecular Structure; Particle Size; Photochemistry; Pyrazines - chemistry; Spectrophotometry; Subcellular Fractions - chemistry; Subcellular Fractions - enzymology; Sucrose - chemistry; Temperature; Watasenia scintillans; BIGCHAP; Bioluminescence; DMSO; Luciferase; Coelenterazine disulfate; TLC; CHAPS; Watasenia scintillans; BSA; SB3-12; MES; DMF; ATP
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/j.bbagen.2008.01.016

The squid Watasenia scintillans emits blue light from numerous photophores. According to Tsuji [F.I. Tsuji, Bioluminescence reaction catalyzed by membrane-bound luciferase in the “firefly squid”, Watasenia scintillans, Biochim. Biophys. Acta 1564 (2002) 189–197.], the luminescence from arm light organs is caused by an ATP-dependent reaction involving Mg 2+, coelenterazine disulfate (luciferin), and an unstable membrane-bound luciferase. We stabilized and partially purified the luciferase in the presence of high concentrations of sucrose, and obtained it as particulates (average size 0.6–2 µm). The ATP-dependent luminescence reaction of coelenterazine disulfate catalyzed by the particulate luciferase was investigated in detail. Optimum temperature of the luminescence reaction is about 5 °C. Coelenterazine disulfate is a strictly specific substrate in this luminescence system; any modification of its structure resulted in a very heavy loss in its light emission capability. The light emitter is the excited state of the amide anion form of coelenteramide disulfate. The quantum yield of coelenterazine disulfate is calculated at 0.36. ATP could be replaced by ATP-γ-S, but not by any other analogues tested. The amount of AMP produced in the luminescence reaction was much smaller than that of coelenteramide disulfate, suggesting that the reaction mechanism of the Watasenia bioluminescence does not involve the formation of adenyl luciferin as an intermediate.