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Such a model was proposed by Ward and Cormier (l976a,b), who later demonstrated that Renilla luciferase and GFP do form a transient, but highly specific, one-to-one heterocomplex under conditions of energy transfer (Ward and Cormier, 1978a). 4 x 10- 6 M GFP on G-l00 Sephadex elutes from the column as a one-to-one complex with GFP (with an apparent molecular weight for the complex of 86,(00); (b) chemical modification of the amino groups of GFP reversibly abolished the energy transfer function without affecting the fluorescence properties of the protein (Table 5); and (c) substitution of either GFP or luciferase, in the in vitro energy transfer reaction mixture, by the homologous protein from another species of coelenterate leads to full energy transfer if both species are from the same genus or from closely related genera (Table 6).

Endo et al. (1970) suggested that this fluorescent steroid is involved in bioluminescence emission. Adam et al. (1977) strengthened this suggestion by showing that protonated ergostatetraenone is an efficient quencher of excited singlet acetone in a model dioxetane chemiluminescence energy transfer system. 1) leads to sensitized chemiluminescence (Amax = 530 nm) in the presence of ergostatetraenone. However, as with the Latia system, the chemical origin of an excited-state donor in fungal luminescence is still open to speculation.

13. A model for in vitro radiationless energy transfer in Renilla bioluminescence. Ward and Cormier (1978a) also reported a marked decrease in assay sensitivity with increasing ionic strength. As the salt concentration was increased from 10 to 100 mM NaCl, the energy transfer efficiency (% green pathway) decreased by nearly a factor of 10. This ionic strength effect may explain why Wampler et al. (1971), who routinely used 100 mM sodium phosphate buffer, required higher GFP concentrations to detect energy transfer.

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