Spectroscopic characterization of the Stentor photoreceptor.
1. On the basis of chromatographic and spectroscopic (absorption, fluorescence and its polarization, fluorescence lifetime, circular dichroism) characterization of the Stentor photoreceptor (stentorin) for photophobic response, the photoreceptor chromophore released from mild acid hydrolysis has been identified as hypericin. 2. The native chromophore is apparently linked to a protein (65 K) containing Lys and several hydrophobic residues, which is soluble in acetone and n-pentane. The peptide-linked stentorin (I) chromophore exhibits circular dichroism in the visible region due to the induced optical activity provided by the peptide. 3. The sodium dodecyl sulfate polyacrylamide gel electrophoresis of a 38% fraction of the sucrose density centrifugation has resolved stentorin II proteins having molecular weights of 13 000, 16 000, 65 000 and 130 000. These proteins, as well as the acetone-soluble peptide, have been spectroscopically characterized with particular emphasis on their primary photoreactivity as the photophobic receptor of Stentor coeruleus. 4. Irradiation of whole living Stentor in dilute buffer solutions induces a decrease in the pH of the medium. A strong dependence upon pH in the fluorescence spectra of both synthetic and native chromophores is also evident, showing a significant drop in the pKa of one or more hydroxyl groups in the excited state. A mechanism for the photophobic response, based on this lowering of the pKa as the primary photoprocess, has been discussed.
D000818:Animals; D000873:Anthracenes; D001551:Benz(a)Anthracenes / Q000302:isolation & purification*; D016798:Ciliophora / Q000032:analysis* / Q000528:radiation effects; D002942:Circular Dichroism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008027:Light; D010569:Perylene / Q000031:analogs & derivatives / Q000302:isolation & purification*; D010616:Phenanthrenes; D010786:Photoreceptor Cells / Q000032:analysis*; D010860:Pigments, Biological / Q000302:isolation & purification*; D011083:Polycyclic Compounds / Q000302:isolation & purification*; D013050:Spectrometry, Fluorescence
D000873:Anthracenes; D001551:Benz(a)Anthracenes; D010616:Phenanthrenes; D010860:Pigments, Biological; D011083:Polycyclic Compounds; C021252:stentorin I; C021253:stentorin II; D010569:Perylene; C004965:hypericin