ISOLATION AND CHARACTERIZATION OF THE PRESUMED PHOTORECEPTOR PROTEIN OF <i>Blepharisma japonicum</i>

Gioffrè, Domenico; Ghetti, Francesco; Lenci, Francesco; Paradiso, C.; Dai, Renke; Song, Pill‐Soon

doi:10.1111/j.1751-1097.1993.tb09561.x

Cited by 33 publications

(18 citation statements)

References 15 publications

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“…3, right series of lanes). Such a low quantity of proteins in these fractions (in the order of nanograms, based on sensitivity of protein silver staining) is consistent with the observed ratio of 280:600 nm absorbance in the chromatography elution profile of this material, characteristic of the purified chromophore (8,9).…”

Section: Resultssupporting

confidence: 82%

“…Both Matsuoka et al (8) and Gioffré et al (9) reported the absorbance spectrum of blepharismin in an organic solvent, and the absorbance spectrum of a purified cell extract in water (in the presence of a detergent) containing the putative chromophore apoprotein of B. japonicum. The spectrum reported in Matsuoka et al (8) shows a remarkable scattering component; max of the chromophore was found to be conserved, and the A 280 /A 588 ratio slightly increased (9). This is not in favor with the presence of a chromophore-apoprotein adduct since the hypothetical protein should have a low A 280 , as compared with the pigment, to maintain the A 280 /A 588 ratio.…”

Section: Introductionmentioning

confidence: 93%

“…This evidence suggested a role for blepharismin as the molecule responsible for photoreception in Blepharisma and led to efforts to identify and purify a protein bearing blepharismin as the *To whom correspondence should be addressed at: Department of Veterinary Anatomy, Biochemistry and Physiology, 2 Viale delle Piagge, 56124 Pisa, Italy. Fax: 39-050-570117; e-mail: apodesta@vet.unipi.it prosthetic group and being responsible for the amplification of the light signal (8,9). Therefore, the purification procedures addressed the isolation of biological material, which absorbs light at wavelengths characteristic both of proteins (280 nm) and of either the photo-oxidized or reduced forms of blepharismin (600 or 580 nm, respectively).…”

Section: Introductionmentioning

confidence: 99%

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Immunological and Biochemical Evidence that Blepharismin is not a Prosthetic Group

Podestà

Gioffrè

Grossi

et al. 2007

Photochemistry and Photobiology

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A polyclonal, multispecific antiserum was raised against a whole 3[(3‐cholamidopropyl)‐dimethylammonio]‐1‐propanesulfonate‐extract of nonirradiated Blepharisma japonicum cells. It was used to reveal the composition of solutions that were hypothesized to contain the photoreceptor of the ciliate. A Bio‐Gel A 1.5 m fine column chromatography of the extract allowed recovery of a single elution peak isolated by recording the 580 nm light absorbance. Fused‐rocket immunoelectrophoresis of this material revealed a large number of >300 kDa coeluted proteins. Blepharismin‐rich material with a molecular mass of approximately 50 kDa, consisting of at least nine proteins was obtained when the same extract underwent preparative isoelectric focusing before column chromatography separation. Purification of the pigment obtained from light‐exposed cells gave blepharismin‐rich material with a molecular weight of approximately 200 kDa. Comparison of the materials obtained under the same conditions, either from the dark‐kept or light‐irradiated cells, by means of pore‐gradient electrophoresis confirmed that proteins present in the two preparations were different. It revealed only a very small amount, if any, of proteins in the chromatography fractions with the highest absorbance at 600 nm. Results are discussed on the basis of the hypothesis that a specific blepharismin‐binding protein does not exist in the protozoan.

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Section: Resultssupporting

confidence: 82%

Section: Introductionmentioning

confidence: 93%

Section: Introductionmentioning

confidence: 99%

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Immunological and Biochemical Evidence that Blepharismin is not a Prosthetic Group

Podestà

Gioffrè

Grossi

et al. 2007

Photochemistry and Photobiology

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“…The data were analyzed by means of the freeware software Imagej (rsb.info.nih.gov/ij), originally developed by NIH (Bethesda, MD, USA). F. salina, B. japonicum, and S. coeruleus were grown as described elsewhere (Marangoni et al, 1996;Gioffré et al, 1993;Walker et al, 1979). Blue B. japonicum was obtained by exposing cells to white light of about 10 W/m 2 for about 24 h.…”

Section: Methodsmentioning

confidence: 99%

Evidence for ciliary pigment localization in colored ciliates and implications for their photosensory transduction chain: A confocal microscopy study

Colombetti¹,

Checcucci²,

Lucia³

et al. 2007

Microscopy Res & Technique

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In this study we report for the first time the localization of a photoreceptor pigment in the cilia of the colored heterotrich ciliates Blepharisma japonicum red and blue form, Fabrea salina, and Stentor coeruleus, as result of a confocal microscopy investigation. Optical sectioning confocal microscopy has been used for studying the spatial distribution of the pigment in the cell body, surprisingly showing that, besides its expected presence in the cortical region immediately below the cell membrane, it is located in the cilia too. In order to ascertain possible differences in the pigment fluorescence properties along the cell body, we have measured emission spectra from different parts of it (anterior, posterior, and cilia). Our results clearly indicate that in all cases the spectra are the same, within experimental errors. Finally, we have evaluated the pigment relative fluorescence efficiency of these ciliates. In an ordered scale from lower to greater efficiency, we have S. coeruleus, B. japonicum blue, B. japonicum red, and F. salina. The possible implications of our findings for the process of photosensory transduction are discussed.

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“…The blepharismin pigment has been reported to be either non-covalently bound to an apoprotein of molecular mass 38 000 dalton (Gioffre et al, 1993;Yamazaki et al, 1993b) or covalently bound to a molecular weight 200,000 protein (Matsuoka et al, 1993). Fabczak et al (2001) reported that phosphorylation of two other proteins, 28and 46-kDa proteins, of B. japonicum is down-and up-regulated by light, respectively.…”

Section: Ciliate Photoreceptorsmentioning

confidence: 99%

Hypericin‐Like Photoreceptors

Song

2005

Handbook of Photosensory Receptors

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The hypericin-derived pigments, stentorin and blepharismin, are the photosensory receptors for the photomotile responses of ciliates Stentor coeruleus and Blepharisma japonicum, respectively. Their photochemistry and its functional linkage to the photosensory signal transduction chain are not well established. In the light signal transduction of the giant single-cell ciliates, a light signal (intensity/wavelength) appears to be amplified by triggering Ca 2+ ion inf lux into the cell. The photosensory signaltransduction pathways of the ciliates include the generation of membrane potentials. The heterotrimeric G-protein(s) may play a signal-transducing role. The photosensory signal transduction for the photophobic response of Stentor coeruleus is represented in the following scheme:Stentorin → G-protein → phosphodiesterase → cGMP/GMP-gated Ca 2+ -channel → ciliary stroke reversal (photophobic response) A similar scheme is likely for the closely related ciliate, Blepharisma japonicum. 20.1

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ISOLATION AND CHARACTERIZATION OF THE PRESUMED PHOTORECEPTOR PROTEIN OF Blepharisma japonicum

Cited by 33 publications

References 15 publications

Immunological and Biochemical Evidence that Blepharismin is not a Prosthetic Group

Immunological and Biochemical Evidence that Blepharismin is not a Prosthetic Group

Evidence for ciliary pigment localization in colored ciliates and implications for their photosensory transduction chain: A confocal microscopy study

Hypericin‐Like Photoreceptors

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