Initial spectroscopic characterization of the ciliate photoreceptor stentorin.

Stentorin serves as the primary photosensor in the single cell ciliate, Stentor coeruleus, for its photophobic and phototactic response to light of visible wavelengths. We separated two subunits, stentorin-2A and -2B, from the previous stentorin complex ('stentorin-2') of greater than half a million molecular mass isolated from the photoreceptor organelle (pigment granule). Stentorin-2B bears the chromophore covalently linked to an approx. 50 kDa apoprotein, as determined by SDS-urea-PAGE. Partial amino acid sequences were obtained from this 50 kDa subunit. Its visible and CD spectra were found to be similar to those of stentorin-2. The steady-state and time-resolved fluorescence spectra of stentorin-2B, in H2O and D2O buffers, were also similar to those of stentorin-2. This suggests that the 50 kDa subunit retains the spectral integrity and primary photoreactivity of the stentorin-complex. The picosecond time-resolved fluorescence study revealed that the short picosecond emission component (tau F approximately equal to 8-10 ps) was the predominant emitting species in stentorin-2B and -2, followed by longer decaying species. No deuterium solvent effect was seen in this fast-decaying species. The possible mechanism for the primary photoreaction appears to involve electron transfer coupled with proton transfer.