Abstract
Polyclonal antibodies raised in rabbits to a mixture of sodium-dodecyl-sulphate-denatured C- and allo-phycocyanin, isolated from Anabaena cylindrica, cross-react with 124-kilodalton (kDa) phytochrome from etiolated oats, in enzyme-linked immunosorbent assays and on Western blots. The component(s) of the anti-phycocyanin serum that cross-reacts with phytochrome appears to be specific for the red-absorbing form of phytochrome (Pr). These antibodies can be detached from Pr by irradiation with red light, and thus show photoreversible binding. This property has been used to immunopurify the anti-phytochrome component from the antiserum using red light as the eluting agent. Competition assays and epitope-mapping studies indicate that the anti-phytochrome component may bind to a site located between 6 and 10 kDa from the amino-terminus of etiolated oat phytochrome.
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Abbreviations
- ELISA:
-
enzyme-linked immunosorbent assay
- kDa:
-
kilodaton
- FR:
-
far-red light
- Pfr:
-
far-red-light-absorbing form of phytochrome
- Pr:
-
red-light-absorbing form of phytochrome
- R:
-
red light
- SDS:
-
sodium dodecyl sulphate
References
Berns, D.S. (1967) Immunochemistry of biliproteins. Plant Physiol. 42, 1569–1586
Bogorad, L. (1975) Phycobiliproteins and complementary chromatic adaptation. Annu. Rev. Plant Physiol. 26, 369–401
Cordonnier, M-M., Greppin, H., Pratt, L.H. (1985) Monoclonal antibodies with differing affinities to the red-absorbing and far-red absorbing forms of phytochrome. Biochemistry 24, 3246–3253
Cundiff, S.L., Pratt, L.H. (1975) Phytochrome characterization by rabbit antiserum against high-molecular weight phytochrome. Plant Physiol. 55, 207–211
Grimm, R., Lottspeich, F., Schneider, H.A.W., Rudiger, W. (1986) Investigations of the peptide chain of 124 kDa phytochrome. Localisation of proteolytic fragments and epitopes for monoclonal antibodies. Z. Naturforsch. 41c, 988–992
Holdsworth, M.L. (1987) Characterization of phytochrome using monoclonal antibodies. Ph.D. Thesis, University of Leicester, UK
Holdsworth, M.L., Whitelam, G.C. (1987) A monoclonal antibody specific for the red-absorbing form of phytochrome. Planta 172, 539–547
Hopkins, D.W., Butler, W.L. (1970) Immunochemical and spectroscopic evidence for protein conformational changes in phytochrome transformation. Plant Physiol. 45, 567–570
Jones, A.M., Vierstra, R.D., Daniels, S.M., Quail, P.H. (1985) The role of separate molecular domains in the structure of phytochrome from etiolated Avena sativa L. Planta 164, 501–506
Kelly, J.M., Lagarias, J.C. (1985) Photochemistry of 124 kilodalton Avena phytochrome under constant illumination invitro. Biochemistry 24, 6003–6010
Pratt, L.H. (1984) Phytochrome purification In: Techniques in photomorphogenesis, pp. 175–200, Smith, H., Holmes, M.G., eds. Academic Press, London
Rice, H.V., Briggs, W.R. (1973) Immunochemistry of phytochrome. Plant Physiol. 51, 939–945
Shimazaki, Y., Cordonnier, MM., Pratt, L.H. (1986) Identification with monoclonal antibodies of a second antigenic domain in Avena phytochrome that changes upon its photoconversion. Plant Physiol. 82, 109–113
Shropshire, W., Mohr, H. (1985) eds. Encyclopedia of plant physiology, N.S. vol. 16A, B: Photomorphogenesis, Springer, Berlin New York
Siegelman, H.W., Turner, B.C., Hendricks, S.B. (1966) The chromophore of phytochrome. Plant Physiol. 41, 1289–1293
Schneider-Poetsch, H.A.W., Schwartz, H., Grimm, R., Rudiger, W. (1988) Cross reactivity of monoclonal antibodies against phytochrome from Zea and Avena. Localisation of epitopes, and an epitope common to monocotyledons, dicotyledons, ferns, mosses and a liverwort. Planta 173, 61–72
Thomas, B., Butcher, G.W., Galfre, G. (1984) Discrimination between the red- and far-red-absorbing forms of phytochrome from Avena sativa L. by monoclonal antibodies. Planta 160, 382–384
Thomas, B., Penn, S.E. (1986) Monoclonal antibody ARC MAC 50.1 binds to a site on the phytochrome molecule which undergoes a photoreversible conformational change. FEBS Lett. 195, 174–178
Vierstra, R.D., Quail, P.H. (1983) Purification and initial characterization of 124 kilodalton phytochrome from Avena. Biochemistry 22, 2498–2505
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Keiller, D.R., Whitelam, G.C. & Smith, H. Polyclonal antibodies raised to phycocyanins contain components specific for the red-absorbing form of phytochrome. Planta 176, 391–398 (1988). https://doi.org/10.1007/BF00395420
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DOI: https://doi.org/10.1007/BF00395420