Abstract
Antibodies raised against two pea (Pisum sativum L. cv. Birte) seed lipoxygenases have been used to analyze lipoxygenase heterogeneity in seeds and in other organs. At least seven different polypeptides were identified in vivo; five of these were identified as precursors synthesized in vitro. The developmental appearance of the seed polypeptides has been analyzed and ‘early’ and ‘late’ forms were identified. Limited N-terminal sequence data indicated further heterogeneity when compared with sequences predicted from cDNAs.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Abbreviations
- cDNA:
-
complementary DNA
- DAF:
-
days after flowering
- HPLC:
-
high-performance liquid chromatography
- Ig:
-
immunoglobulin
- kb:
-
kilobase
- Mr :
-
relative molecular mass
- PAGE:
-
polyacrylamide gel electrophoresis
- PVDF:
-
polyvinylidene difluoride
- SDS:
-
sodium dodecyl sulphate
- SSC:
-
0.15 M sodium chloride, 0.015 M sodium citrate, pH 7.0
References
Ambrose, M.J., Wang, T.L., Cook, S.K., Hedley, C.L. (1987) An analysis of seed development in Pisum sativum. IV. Cotyledon cell populations in vivo and in vitro. J. Exp. Bot. 38, 1909–1920
Anstis, P.J.P., Friend, J. (1974a) The effect of light on lipoxygenase activity in dwarf pea seedlings. Phytochemistry 13, 2709–2712
Anstis, P.J.P., Friend, J. (1974b) The isoenzyme distribution of etiolated pea seedling lipoxygenase. Planta 115, 329–335
Arens, D., Seilmeier, W., Weber, F., Kloos, G., Grosch, W. (1973) Purification and properties of a carotene co-oxidizing lipoxy-genase from peas. Biochim. Biophys. Acta 327, 295–305
Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254
Casey, R. (1979) Immunoaffinity chromatography as a means of purifying legumin from Pisum (pea) seeds. Biochem. J. 177, 509–520
Casey, R., Domoney, C., Nielsen, N.C. (1985) Isolation of a cDNA clone for pea (Pisum sativum) seed lipoxygenase. Biochem. J. 232, 79–85
Chandler, P.M., Spencer, D., Randall, P.J., Higgins, T.J.V. (1984) Influence of sulfur nutrition on developmental patterns of some major pea seed proteins and their mRNAs. Plant Physiol. 75, 651–657
Domoney, C., Casey, R. (1983) Cloning and characterization of complementary DNA for convicilin, a major seed storage protein in Pisum sativum L. Planta 159, 446–453
Domoney, C., Casey, R. (1985) Measurement of gene number for seed storage proteins in Pisum. Nucleic Acids Res. 13, 687–699
Domoney, C., Casey, R. (1987) Changes in legumin messenger RNAs throughout seed development in Pisum sativum L. Planta 170, 562–566
Dixon, R.A.F., Jones, R.E., Diehl, R.E., Bennett, C.D., Kargman, S., Rouzer, C.A. (1988) Cloning of the cDNA for human 5-lipoxygenase. Proc. Natl. Acad. Sci. USA 85, 416–420
Ealing, P.M., Casey, R. (1988) The complete amino acid sequence of a pea (Pisum sativum) seed lipoxygenase predicted from a near full-length cDNA. Biochem. J. 253, 915–918
Eriksson, C.E., Svensson, S.G. (1970) Lipoxygenase from peas, purification and properties of the enzyme. Biochim. Biophys. Acta 198, 449–459
Galliard, T., Chan, H.W.-S. (1980) Lipoxygenases. In: The biochemistry of plants, vol. 4, pp. 131–161, Stumpf, P.K., ed. Academic Press, New York
Gatehouse, J.A., Bown, D., Gilroy, J., Levasseur, M., Castleton, J., Ellis, T.H.N. (1988) Two genes encoding ‘minor’ legumin polypeptides in pea (Pisum sativum L.). Characterization and complete sequence of the LegJ gene. Biochem. J. 250, 15–24
Haydar, M., Steele, L., Hadziyev, D. (1975) Oxidation of pea lipids by pea seed lipoxygenase. J. Food Sci. 40, 808–814
Hedley, C.L., Smith, C.M., Ambrose, M.J., Cook, S.K., Wang, T.L. (1986) An analysis of seed development in Pisum sativum. II. The effect of the r-locus on the growth and development of the seed. Ann. Bot. 58, 371–379
Hildebrand, D.F. (1989) Lipoxygenases. Physiol. Plant. 76, 249–253
Kerckaert, J.-P., Bayard, B., Debray, H., Sautière, P., Biserte, G. (1977) Rat alpha-fetoprotein heterogeneity. Comparative chemical study of the two electrophoretic variants and their Ricinus lectin-binding properties. Biochim. Biophys. Acta 493, 293–303
Kühn, H., Heydeck, D., Wiesner, R., Schewe, T. (1985) The positional specificity of wheat lipoxygenase; the carboxylic group as signal for the recognition of the site of the hydrogen removal. Biochim. Biophys. Acta 830, 25–29
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685
Matsudaira, P. (1987) Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J. Biol. Chem. 262, 10035–10038
Matsumoto, T., Funk, C.D., Rådmark, O., Höög, J.-O., Jörnvall, H., Samuelsson, B. (1988) Molecular cloning and amino acid sequence of human 5-lipoxygenase. Proc. Natl. Acad. Sci. USA 85, 26–30
North, H., Casey, R., Domoney, C. (1989) Inheritance and mapping of seed lipoxygenase polypeptides in Pisum. Theor. Appl. Genet. 77, 805–808
O'Farrell, P.H. (1975) High resolution two-dimensional electrophoresis of proteins. J. Biol. Chem. 250, 4007–4021
Ramadoss, C.S., Axelrod, B. (1982) High-performance liquid Chromatographic separation of lipoxygenase isozymes in crude soybean extracts. Anal. Biochem. 127, 25–31
Reddanna, P., Whelan, J., Reddy, P.S., Reddy, C.C. (1988) Isolation and characterization of 5-lipoxygenase from tulip bulbs. Biochem. Biophys. Res. Comm. 157, 1348–1351
Shibata, D., Steczko, J., Dixon, J.E., Hermodson, M., Yazdanparast, R., Axelrod, B. (1987) Primary structure of soybean lipoxygenase-1. J. Biol. Chem. 262, 10080–10085
Shibata, D., Steczko, J., Dixon, J.E., Andrews, P.C., Hermodson, M., Axelrod, B. (1988) Primary structure of soybean lipoxygenase L-2. J. Biol. Chem. 263, 6816–6821
Shimizu, T., Rådmark, O., Samuelsson, B. (1984) Enzyme with dual lipoxygenase activities catalyzes leukotriene A4 synthesis from arachidonic acid. Proc. Natl. Acad. Sci. USA 81, 689–693
Sigal, E., Craik, C.S., Highland, E., Grunberger, D., Costello, L.L., Dixon, R.A.F., Nadel, J.A. (1988) Molecular cloning and primary structure of human 15-lipoxygenase. Biochem. Biophys. Res. Comm. 157, 457–464
Spaapen, L.J.M., Vliegenthart, J.F.G., Boldingh, J. (1977) On the properties of a pea lipoxygenase. Biochim. Biophys. Acta. 488, 517–520
Vernooy-Gerritsen, M., Bos, A.L.M., Veldink, G.A., Vliegenthart, J.F.G. (1983) Localization of lipoxygenases-1 and -2 in germinating soybean seeds by an indirect immunofluorescence technique. Plant Physiol. 73, 262–267
Yenofsky, R.L., Fine, M., Liu, C. (1988) Isolation and characterization of a soybean (Glycine max) lipoxygenase-3 gene. Mol. Gen. Genet. 211, 215–222
Yoon, S., Klein, B.P. (1979) Some properties of pea lipoxygenase isoenzymes. J. Agric. Food Chem. 27, 955–962
Yuen, S., Hunkapiller, M.W., Wilson, K.J., Yuan, P.M. (1986) SDS-PAGE electroblotting. Applied Biosystems User Bulletin, Issue no. 25
Author information
Authors and Affiliations
Additional information
This work was supported by the Agricultural and Food Research Council via a grant-in-aid to the John Innes Institute. We acknowledge financial support from the Commission of the European Communities Biotechnology Action Programme; grant No. 0063-UK.
Rights and permissions
About this article
Cite this article
Domoney, C., Firmin, J.L., Sidebottom, C. et al. Lipoxygenase heterogeneity in Pisum sativum . Planta 181, 35–43 (1990). https://doi.org/10.1007/BF00202322
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00202322