Abstract
The hypothesis was tested that reversible riboflavin (RF)-binding sites are part of the photoreceptor in Euglena gracilis. Published evidence shows that the phototactic stimulus — with a flavin-type action spectrum — is perceived at the paraflagellar body (PFB). Flagella with PFBs were isolated from Euglena gracilis by a combined cold and Ca2+ shock. Saturable binding of [14C]RF was demonstrated with such preparations, in the oxidized state as well as under reducing conditions in the presence of dithionite. Affinities for RF were high: K D (oxidized)=0.08 μM, and K D (reduced)=0.7 μM. Flavin mononucleotide and flavin adenine dinucleotide showed lower binding affinities. The in vitro RF binding per unit of protein was enriched approximately tenfold in the flagellar preparations when compared with homogenates of whole cells. The number of (reduced) binding sites per entire flagellum was determined to be in the order of 106. This number is in line with published estimates of chromophores bound in or at the PFB.
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Abbreviations
- FAD:
-
flavin adenine dinucleotide
- FMN:
-
flavin mononucleotide
- PFB:
-
paraflagellar body
- RF:
-
riboflavin
References
Beale, S.I., Foley, T., Dzelzkalns, V. (1981) δ-Aminolevulinic acid synthase from Euglena gracilis. Proc. Natl. Acad. Sci. USA 78, 1666–1669
Benedetti, P.A., Checcucci, A. (1975) Paraflagellar body (PFB) pigments studied by fluorescence microscopy in Euglena gracilis. Plant Sci. Letts. 4, 47–51
Brodhun, B., Häder, D.P. (1990) Photoreceptor proteins and pigments in the paraflagellar body of the flagellate Euglena gracilis. Photochem. Photobiol. 52, 865–872
Checcucci, A., Colombetti, G., Ferrara, R., Lenci, F. (1976) Action spectra for photoaccumulation of green and colorless Euglena: evidence for identification of receptor pigments. Photochem. Photobiol. 23, 51–54
Coleman, A.W. (1988) The autofluorescent flagellum: a new phylogenetic enigma. J. Phycol. 24, 118–120
Colombetti, G., Lenci, F. (1980) Identification and spectroscopic characterization of photoreceptor pigments. In: Photoreception and sensory transduction in aneural organisms (NATO ASI-Series A 33), pp. 173–188, Lenci, F., Colombetti, G., eds. Plenum Press, New York
Colombetti, G., Lenci, F., Diehn, B. (1982) Responses to photic, chemical, and mechanical stimuli. In: The biology of Euglena III. pp. 169–196, Buetow, D.E., ed. Academic Press, New York
Dohrmann, U. (1983) In vitro riboflavin binding and endogenous flavins in Phycomyces blakesleeanus. Planta 159, 357–365
Fritz, B.J., Kasai, S., Matsui, K. (1989) Free cellular riboflavin is involved in phase shifting by light of the circadian clock in Neurospora crassa. Plant Cell Physiol. 30, 557–564
Galland, P., Senger, H. (1988) New trends in photobiology: The role of flavins as photoreceptors. J. Photochem. Photobiol. B 1, 277–294
Galland, P., Keiner, P., Dörnemann, D., Senger, H., Brodhun, B., Häder, D.P. (1990) Pterin- and flavin-like fluorescence associated with isolated flagella of Euglena gracilis. Photochem. Photobiol. 51, 675–680
Gibbs, S.P. (1978) The chloroplasts of Euglena may have evolved from symbiotic green algae. Can. J. Bot. 56, 2883–2889
Gualtieri, P., Barsanti, L., Rosati, G. (1986) Isolation of the photoreceptor (paraflagellar body) of the phototactic flagellate Euglena gracilis. Arch. Microbiol. 145, 303–305
Gualtieri, P., Barsanti, L., Passarelli, V. (1989) Absorption spectrum of a single paraflagellar swelling of Euglena gracilis. Biochim. Biophys. Acta 993, 293–296
Harlow, E., Lane, D. (1988) Antibodies: a laboratory manual. Cold Spring Harbor Lab., New York
Hertel, R. (1980) Phototropism in lower plants. In: Photoreception and sensory transduction in aneural organisms (NATO ASI-Series A 33), pp. 89–105, Lenci, F., Colombetti, G., eds. Plenum Press, New York
Hertel, R., Jesaitis, A.J., Dohrmann, U., Briggs, W.R. (1980) In vitro binding of riboflavin to subcellular particles from maize coleoptiles and Cucurbita hypocotyls. Planta 147, 312–319
Kawai, H (1988) A flavin-like autofluorescent substance in the posterior flagellum of golden and brown algae. J. Phycol. 24, 114–117
Kuznicki, L., Mikolajzcyk, E., Walne, P.L. (1990) Photobehavior of euglenoid flagellates: theoretical and evolutionary perspectives. Crit. Rev. Plant Sci. 9, 343–369
Laemmli, U.K. (1970) Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227, 680–685
Massey, V., Palmer, G. (1966) On the existence of spectrally distinct classes of flavoprotein semiquinones. Biochemistry 5, 3181–3189
Müller, F., Massey, V. (1969) Flavin-sulfite complexes and their structures. J. Biol. Chem. 224, 4007–4016
Piccini, E., Mammi, M. (1978) Motor apparatus of Euglena gracilis: ultrastructure of the basal portion of the flagellum and the paraflagellar body. Boll. Zool. 45, 405–414
Presti, D.E., Hsu, W.J., Delbrück, M. (1977) Phototropism in Phycomyces mutants lacking β-carotene. Photochem. Photobiol. 26, 403–405
Schlösser, U.G. (1982) Sammlung von Algenkulturen. Ber. Dtsch. Bot. Ges. 95, 181–276
Schmidt, W., Galland, P., Senger, H., Furuya, M. (1990) Micro-spectrophotometry of Euglena gracilis. Pterin- and flavin-like fluorescence in the paraflagellar body. Planta 182, 374–381
Spector, T. (1978) Refinement of the Coomassie-Blue-method of protein quantitation. Annal. Biochem. 86, 142–146
Thein, M., Michalke, W. (1988) Bisulfite interacts specifically with binding sites of the auxin transport inhibitor N-1-Naphthylphthalamic acid. Planta 76, 343–350
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This work was supported by the Deutsche Forscungsgemeinschaft.
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Nebenführ, A., Schäfer, A., Galland, P. et al. Riboflavin-binding sites associated with flagella of Euglena: A candidate for blue-light photoreceptor?. Planta 185, 65–71 (1991). https://doi.org/10.1007/BF00194516
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DOI: https://doi.org/10.1007/BF00194516