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Changes in lipoxygenase isoforms during germination and early seedling growth of Pisum sativum L.

Published online by Cambridge University Press:  22 February 2007

Bing Mo  and
Karen L. Koster
Bing Mo
Affiliation:
Department of Biology, University of South Dakota, Vermillion, SD 57069, USA
Karen L. Koster*
Affiliation:
Department of Biology, University of South Dakota, Vermillion, SD 57069, USA
*
*Correspondence: Fax: +1-605-677-6557 Email: Karen.Koster@usd.edu
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Abstract

Investigations of physiological functions of seed lipoxygenase during germination and early seedling growth are complicated by the presence of multiple isoforms with different product specificity. The combination of isoelectric focusing, lipoxygenase in-gel activity staining and mass spectrometry showed that only two seed lipoxygenases, LOX-2 and LOX-3, were active at neutral pH in protein extracts from dry and germinating pea seeds (Pisum sativum L.), with LOX-3 more abundant than LOX-2. Each active seed LOX appeared at multiple pIs, probably resulting from post-translational modifications. The abundance of pea seed lipoxygenases was lower in embryonic shoots than in other parts of the axis. After radicle emergence, seed LOXs were degraded, as revealed by Western blotting using antibodies specific to pea seed LOXs. Two new vegetative isoforms, LOX-N3 and LOX-g, appeared in the shoot and gradually replaced the pea seed lipoxygenases as the principal contributors to LOX enzyme activity during early seedling growth. Activities of LOX-N3 and LOX-g were abundant in shoot tips, but not root tips, in pea seedlings. Thus, during germination, there is a shift of LOX activity from radicles to shoots that accompanies the transition from seed LOXs to vegetative LOXs. These results confirmed that identification of pea LOXs based solely on pI may be misleading; however, mass spectrometry enabled more accurate identification of these highly homologous proteins to be made.

Keywords

lipoxygenaseLOXPisum sativumprotein identificationseed germination
Type
Research Article
Information
Seed Science Research , Volume 16 , Issue 2 , June 2006 , pp. 97 - 106
Copyright
Copyright © Cambridge University Press 2006

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References

Andersen, J.S. and Mann, M. (2000) Functional genomics by mass spectrometry. FEBS Letters 480, 25–31.Google Scholar
Anstis, P.J.P. and Friend, J. (1974) The isoenzyme distribution of etiolated pea seedling lipoxygenase. Planta 115, 329–335.Google Scholar
Bell, E. and Mullet, J.E. (1991) Lipoxygenase gene expression is modulated in plants by water deficit, wounding, and methyl jasmonate. Molecular and General Genetics 230, 456–462.Google Scholar
Blom, N., Gammeltoft, S. and Brunak, S. (1999) Sequence- and structure-based prediction of eukaryotic protein phosphorylation sites. Journal of Molecular Biology 294, 1351–1362.Google Scholar
Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248–254.Google Scholar
Burghoffer, C., Chardot, T. and Meunier, J.C. (1999) The phosphorylation and characterisation of soybean isolate and its β-conglycinin component by casein kinase II. Journal of the Science of Food and Agriculture 79, 1179–1182.Google Scholar
Chateigner, A.L. Le, Deunff, Y. and Jalouzot, R. (1999) Germination-associated changes in the transcript content of pea seedling lipoxygenases. Lipoxygenase-g: a new marker of axis growth resumption. Planta 208, 606–613.CrossRefGoogle Scholar
Domoney, C., Firmin, J.L., Sidebottom, C., Ealing, P.M., Slabas, A. and Casey, R. (1990) Lipoxygenase heterogeneity in Pisum sativum. Planta 181, 35–43.Google Scholar
Feussner, I., Kühn, H. and Wasternack, C. (2001) Lipoxygenase-dependent degradation of storage lipids. Trends in Plant Science 6, 268–273.Google Scholar
Forster, C., North, H., Afzal, N., Domoney, C., Hornostaj, A., Robinson, D.S. and Casey, R. (1999) Molecular analysis of a null mutant for pea (Pisum sativum L.) seed lipoxygenase-2. Plant Molecular Biology 39, 1209–1220.CrossRefGoogle Scholar
Gobel, C., Feussner, I., Schmidt, A., Scheel, D., Sanchez-Serrano, J., Hamberg, M. and Rosahl, S. (2001) Oxylipin profiling reveals the preferential stimulation of the 9-lipoxygenase pathway in elicitor-treated potato cells. Journal of Biological Chemistry 276, 6267–6273.CrossRefGoogle ScholarPubMed
Harlow, E. and Lane, D. (1988) Antibodies: A laboratory manual. Cold Spring Harbor, NY, Cold Spring Harbor Laboratory. pp. 421–470.Google Scholar
Herbert, B., Hopwood, F., Oxley, D., McCarthy, J., Laver, M., Grinyer, J., Goodall, A., Williams, K., Castagna, A. and Righetti, P.G. (2003) β-elimination: an unexpected artefact in proteome analysis. Proteomics 3, 826–831.CrossRefGoogle ScholarPubMed
Hildebrand, D.F., Versluys, R.T. and Collins, G.B. (1991) Changes in lipoxygenase isozyme levels during soybean embryo development. Plant Science 75, 1–8.Google Scholar
Holtman, W.L., Roberts, M.R. and Wang, M. (2000) 14-3-3 proteins and a 13-lipoxygenase form associations in a phosphorylation-dependent manner. Biochemical Society Transactions 28, 834–836.CrossRefGoogle Scholar
Hughes, R.K., Wu, Z., Robinson, D.S., Hardy, D., West, S.I., Fairhurst, S.A. and Casey, R. (1998) Characterization of authentic recombinant pea-seed lipoxygenases with distinct properties and reaction mechanisms. Biochemical Journal 333, 33–43.CrossRefGoogle ScholarPubMed
Hughes, R.K., Lawson, D.M., Hornostaj, A.R., Fairhurst, S.A. and Casey, R. (2001) Mutagenesis and modelling of linoleate-binding to pea seed lipoxygenase. European Journal of Biochemistry 268, 1030–1040.CrossRefGoogle ScholarPubMed
Kinter, M. and Sherman, N.E. (2000) Protein sequencing and identification using tandem mass spectrometry. New York, Wiley-Interscience.Google Scholar
Kolomiets, M.V., Hannapel, D.J., Chen, H., Tymeson, M. and Gladon, R.J. (2001) Lipoxygenase is involved in the control of potato tuber development. Plant Cell 13, 613–626.Google Scholar
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.Google Scholar
Loiseau, J., Vu, B.L., Macherel, M.H., Le, Deunff, Y. (2001) Seed lipoxygenases: Occurrence and functions. Seed Science Research 11, 199–211.Google Scholar
Maccarrone, M., van Aarle, P.G.M., Veldink, G.A. and Vliegenthart, J.F.G (1994) In vitro oxygenation of soybean biomembranes by lipoxygenase-2. Biochimica et Biophysica Acta 1190, 164–169.Google Scholar
Maccarrone, M., Salucci, M.L., van Zadelhoff, G., Malatesta, F., Veldink, G., Vliegenthart, J.F.G., Finazzi-Agro, A. (2001) Tryptic digestion of soybean lipoxygenase-1 generates a 60?kDa fragment with improved activity and membrane binding ability. Biochemistry 40, 6819–6827.Google Scholar
Marcantonio, E.E. and Hynes, R.O. (1988) Antibodies to the conserved cytoplasmic domain of the integrin beta 1 subunit react with proteins in vertebrates, invertebrates, and fungi. Journal of Cell Biology 106, 1765–1772.Google Scholar
Mo, B. (2004) Plant ‘integrin-like’ proteins in pea (Pisum sativum L.) embryonic axes. PhD Dissertation, Department of Biology, University of South Dakota.Google Scholar
Mo, B. and Koster, K.L. (2003) An ‘integrin-like’ protein in pea embryos is identified as lipoxygenase Plant Biology 2003 – The Annual Meeting of the American Society of Plant Biologists, Abstract 1362.Google Scholar
Park, T.K. and Polacco, J.C. (1989) Distinct lipoxygenase species appear in the hypocotyls/radicle of germinating soybean. Plant Physiology 90, 285–290.CrossRefGoogle ScholarPubMed
Perkins, D.N., Pappin, D.J.C., Creasy, D.M. and Cottrell, J.S. (1999) Probability-based protein identification by searching sequence databases using mass spectrometry data. Electrophoresis 20, 3551–3567.3.0.CO;2-2>CrossRefGoogle ScholarPubMed
Porta, H., Rocha-Sosa, M. (2002) Plant lipoxygenases. Physiological and molecular features. Plant Physiology 130, 15–21.Google Scholar
Porta, H., Rueda-Benitez, P., Campos, F., Colmenero-Flores, J.M., Colorado, J.M., Carmona, M.J., Covarrubias, A.A., Rocha-Sosa, M. (1999) Analysis of lipoxygenase mRNA accumulation in the common bean (Phaseolus vulgaris L.) during development and under stress conditions. Plant and Cell Physiology 40, 850–858.Google Scholar
Reisdorph, N.A. and Koster, K.L. (1999) Progressive loss of desiccation tolerance in germinating pea (Pisum sativum) seeds. Physiologia Plantarum 105, 266–271.Google Scholar
Robertson, E.F., Dannelly, H.K., Malloy, P.J. and Reeves, H.C. (1987) Rapid isoelectric focusing in a vertical polyacrylamide minigel system. Analytical Biochemistry 167, 290–294.Google Scholar
Rodriguez-Concepción, M., Gómez, M.D., Beltrán, J.P. (1996) Immunolocalization of lipoxygenase in pea (Pisum sativum L.) carpels. Plant Cell Reports 15, 620–626.Google Scholar
Sanz, L.C., Pérez, A.G., Ríos, J.J., Olías, J.M. (1993) Positional specificity of ketodienes from linoleic acid aerobically formed by lipoxygenase isozymes from kidney beans and pea. Journal of Agricultural and Food Chemistry 41, 696–699.CrossRefGoogle Scholar
Saravitz, D.M. and Siedow, J.N. (1995) The lipoxygenase isoforms in soybean [Glycine max (L.) Merr. ] leaves. Plant Physiology 107, 535–543.CrossRefGoogle Scholar
Siedow, J.N. (1991) Plant lipoxygenase: Structure and function. Annual Review of Plant Physiology and Plant Molecular Biology 42, 145–188.Google Scholar
Smith, J.J., Linforth, R. and Tucker, G.A. (1997) Soluble lipoxygenase isoforms from tomato fruit. Phytochemistry 45, 453–458.CrossRefGoogle Scholar
Song, Y., Love, M.H. and Murphy, P. (1990) Subcellular localization of lipoxygenase-1 and -2 in germinating soybean seeds and seedlings. Journal of the American Oil Chemists Society 67, 961–965.Google Scholar
Vollenweider, S., Weber, H., Stolz, S., Chetelat, A. and Farmer, E.E. (2000) Fatty acid ketodienes and fatty acid ketotrienes: Michael addition acceptors that accumulate in wounded and diseased Arabidopsis leaves. Plant Journal 24, 467–476.Google Scholar
Wang, C., Croft, K.P.C., Jarlfors, U. and Hildebrand, D.F. (1999) Subcellular localization studies indicate that lipoxygenases 1 to 6 are not involved in lipid mobilization during soybean germination. Plant Physiology 120, 227–235.Google Scholar
Werz, O. and Steinhilber, D. (2005) Development of 5-lipoxygenase inhibitors – lessons from cellular enzyme regulation. Biochemical Pharmacology 70, 327–333.Google Scholar
Wisniewski, J.P., Gardner, C.D. and Brewin, N.J. (1999) Isolation of lipoxygenase cDNA clones from pea nodule mRNA. Plant Molecular Biology 39, 775–783.Google Scholar
Wu, Z.C., Robinson, D.S., Domoney, C. and Casey, R. (1995) High-performance liquid chromatographic analysis of the products of linoleic acid oxidation catalyzed by pea (Pisum sativum) seed lipoxygenases. Journal of Agricultural and Food Chemistry 43, 337–342.CrossRefGoogle Scholar
Yoon, S. and Klein, B.P. (1979) Some properties of pea lipoxygenase isoenzymes. Journal of Agricultural and Food Chemistry 27, 955–962.Google Scholar