Abstract
An unknown brownish protein was purified by ammonium sulfate precipitation and DEAE-cellulose column and hydroxyapatite column chromatographies from pumpkin callus treated with a high concentration of 2,4-D. The apparent molecular mass and isoelectric point of the purified protein were estimated to be 38 kD and 4.6, respectively. The absorption spectra of the protein showed a shoulder at around 280 nm and a sharp peak at 405 nm. In order to determine what the purified protein is, a cDNA library of the callus treated with a high concentration of 2,4-D was immunoscreened with antiserum raised against the purified protein. The obtained positive cDNA clone encoded a thioredoxin h having a predicted molecular mass of 13 123 D and a predicted isoelectric point of 5.24, suggesting that the purified protein might be a trimer that was formed by oxidative polymerization of the thioredoxin h.
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Bower, M.S., Matias, D.D., Fernandes-Carvalho, E., Mazzurco, M., Gu, T., Rothstein, S.J., Goring, D.R.: Two members of the thioredoxin-h family interact with the kinase domain of a Brassica S locus receptor kinase.-Plant Cell 8: 1641–1650, 1996.
Bradford, M.M.: A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.-Anal. Biochem. 72: 248–254, 1976.
Brugidou, C., Marty, I., Chartier, Y., Meyer, Y.: The Nicotiana tabacum genome encodes two cytoplasmic thioredoxin genes which are differently expressed.-Mol. gen. Genet. 238: 285–293, 1993.
Cabrillac, D., Cock, J.M., Dumas, C., Gaude, T.: The S-locus receptor kinase is inhibited by thioredoxins and activated by pollen coat proteins.-Nature 410: 220–223, 2001.
Crawford, N.A., Droux, M., Kosower, N.S., Buchanan, B.B.: Evidence for function of the ferredoxin thioredoxin system in the reductive activation of target enzymes of isolated intact chloroplasts.-Arch. Biochem. Biophys. 271: 223–239, 1989.
Droux, M., Miginiac-Maslow, M., Jacquot, J.P., Gadal, P., Crawford, N., Kosower, A., Buchanan, B.B.: Ferredoxin-thioredoxin reductase: a catalytically active dithiol group links ferredoxin to thioredoxin functional in photosynthetic enzyme regulation.-Arch. Biochem. Biophys. 256: 372–380, 1987.
Eklund, H., Cambillau, C., Sjoberg, B.M., Holmgren, A., Jornvall, H., Hoog, J.O., Branden, C.I.: Conformational and functional similarities between glutaredoxin and thioredoxins.-EMBO J. 3: 1443–1449, 1984.
Fujita, M., Adachi, Y., Hanada, Y.: Preliminary characterization of glutathione S-transferases that accumulate in callus of pumpkin (Cucurbita maxima Duch.).-Plant Cell Physiol. 35: 275–282, 1994.
Gautier, M.-F., Lullien-Pellerin, V., De Lamotte-Guery, F., Guirao, A., Joudrier, P.: Characterization of wheat thioredoxin h cDNA and production of an active Triticum aestivum protein in Escherichia coli.-Eur. J. Biochem. 252: 314–324, 1998.
Holmgren, A.: Thioredoxin.-Annu. Rev. Biochem. 54: 237–271, 1985.
Ishiwatari, Y., Fujiwara, T., McFarland, K.C., Nemoto, K., Hayashi, H., Chino, M., Lucas, W.J.: Rice phloem thioredoxin h has the capacity to mediate its own cell-to-cell transport through plasmodesmata.-Planta 205: 12–22, 1998.
Ishiwatari, Y., Honda, C., Kawashima, I., Nakamura, S., Hirano, H., Mori, S., Fujiwara, T., Hayashi, H., Chino, M.: Thioredoxin h is one of the major proteins in rice phloem sap.-Planta 195: 456–463, 1995.
Li, X., Nield, J., Hayman, D., Langridge, P.: Thioredoxin activity in the C terminus of Phalaris S protein.-Plant J. 8: 133–138, 1995.
Marty, I., Meyer, Y.: Nucleotide sequence of a cDNA encoding a tobacco thioredoxin.-Plant mol. Biol. 17: 143–147, 1991.
Rey, P., Pruvot, G., Becuwe, N., Eymery, F., Rumeau, D., Peltier, G.: A novel thioredoxin-like protein located in chloroplast is induced by water deficit in Solanum tuberosum L. plants.-Plant J. 13: 97–107, 1998.
Rivera-Madrid, R., Mestres, D., Marinho, P., Jacquot, J.-P., Decottignies, P., Miginiac-Maslow, M., Meyer, Y.: Evidence for five divergent thioredoxin h sequences in Arabidopsis thaliana.-Proc. nat. Acad. Sci. USA 92: 5620–5624, 1995.
Scheibe, R.: Redox-modulation of chloroplast enzymes: a common principle for individual control.-Plant Physiol. 96: 1–3, 1991.
Stein, M., Jacquot, J.-P., Jeannette, E., Decottignies, P., Hodges, M., Lancelin, J.-M., Mittard, V., Schmitter, J.-M., Miginiac-Maslow, M.: Chlamydomonas reinhardtii thioredoxins: structure of the genes coding for the chloroplastic m and cytosolic h isoforms: expression in Escherichia coli of the recombinant proteins, purification and biochemical properties.-Plant mol. Biol. 28: 487–503, 1995.
Toriyama, K., Hanaoka, K., Okada, T., Watanabe, M.: Molecular cloning of a cDNA encoding a pollen extracellular protein as a potential source of a pollen allergen in Brassica rapa.-FEBS Lett. 424: 234–238, 1998.
Vries, S.D., Hoge, H., Bisseling, T.: Isolation of total and polysomal RNA from plant tissues.-Plant mol. Biol. Manual B6: 1–13, 1988.
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Fujita, M., Hossain, M. Brownish Acidic Protein Induced in Pumpkin Callus by a High Concentration of 2,4-Dichlorophenoxyacetic Acid. Biologia Plantarum 46, 175–179 (2003). https://doi.org/10.1023/A:1022877905774
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DOI: https://doi.org/10.1023/A:1022877905774