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Calcium-dependent phospholipid-binding proteins in plants

Their characterisation and potential for regulating cell growth

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Abstract

There is evidence that Ca2+ can regulate vesicle-mediated secretion in plant cells, but the mechanism for this is not known. One possibility is that Ca2+ -dependent phospholipid-binding proteins (annexins) couple the Ca2+ stimulus to the exocytotic response. Using a protocol developed for the isolation of animal annexins we have identified proteins in maize (Zea mays L.) coleoptiles that have similar characteristics to annexins. The predominant polypeptide species run as a doublet of relative molecular mass (Mr) 33000–35000 on sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE); another less-abundant protein of Mr 23000 is also present. In the presence of Ca2+ these proteins bind to liposomes composed of acidic phospholipids. Calcium-sensitivity of binding differs for each protein and is also influenced by the pH of the buffer used for the liposome-binding assay. Antiserum raised to the 33 to 35-kDa doublet purified on SDS-PAGE recognises the doublet in crude extracts from maize and proteins of similar Mr in Tradescantia virginiana and tobacco Nicotiana tabacum L. The antiserum also recognises p68 (Annexin VI) from chicken gizzard extracts, indicating homology between animal annexins and the maize proteins. For the maize proteins to be involved in the regulation of exocytosis, binding to phospholipids would be expected to occur at physiological levels of Ca2+. The characteristics of the maize annexin-like proteins are described and attention drawn to the marked effect of pH in lowering the requirement for Ca2+ for phospholipid binding.

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Abbreviations

DEAE:

diethylaminoethyl

EGTA:

ethylene glycol-bis (β-aminoethyiether)-N,N,N′,N′-tetraacetic acid

kDa:

kilodalton(s)

Mr :

relative molecular mass

SDS-PAGE:

sodium dodecyl sulphate-polyacrylamide gel electrophoresis

References

  • Ali, S.M., Geisow, M.J., Burgoyne, R.D. (1989) A role for calpactin in calcium-dependent exocytosis in adrenal chromaffin cells. Nature 340, 313–315

    Google Scholar 

  • Baydoun, E.A-H., Northcote, D.H. (1980) Isolation and characterisation of membranes from the cells of maize root tips. J. Cell Sci. 45, 147–167

    Google Scholar 

  • Baydoun, E.A-H., Northcote, D.H. (1980) Measurement and characteristics of fusion of isolated membrane fractions from maize root tips. J. Cell Sci. 45, 169–186

    Google Scholar 

  • Baydoun, E.A-H., Northcote, D.H. (1981) The extraction from maize (Zea mays) root cells of membrane-bound protein with Ca2 + -dependent ATPase activity and its possible role in membrane fusion in vitro. Biochem. J. 193, 781–792

    Google Scholar 

  • Boustead, C., Walker, J.H., Geisow, M.J. (1988) Isolation and characterisation of two novel calcium-dependent phospholipidbinding proteins from bovine lung. FEBS Lett. 233, 233–238

    Google Scholar 

  • Boustead, C., Smallwood, M., Small, H., Bowles, D.J., Walker, J.H. (1989) Identification of calcium-dependent phospholipidbinding proteins in higher plant cells. FEBS Lett. 244, 456–460

    Google Scholar 

  • Brummell, D.A., Hall, J.L. (1987) Rapid cellular responses to auxin and the regulation of growth. Plant Cell Environ. 10, 523–543

    Google Scholar 

  • Brummell, D.A., Maclachlan, G.A. (1989) Calcium antagonist TMB-8 inhibits cell wall formation and growth in pea. J. Exp. Bot. 40, 559–565

    Google Scholar 

  • Burgoyne, R.D., Geisow, M.J. (1989) The annexin family of calcium-binding proteins. Cell Calcium 10, 1–10

    Google Scholar 

  • Clarkson, D.T., Brownlee, C., Ayling, S.M. (1988) Cytoplasmic calcium measurements in intact higher plant cells: results from fluorescence ratio imaging of fura-2. J. Cell Sci. 91, 71–80

    Google Scholar 

  • Creutz, C.E., Pazoles, C.J., Pollard, H.B. (1978) Identification and purification of an adrenal medullary protein (synexin) that causes calcium-dependent aggregation of isolated chromaffin granules. J. Biol. Chem. 253, 2858–2866

    Google Scholar 

  • Crompton, M.R., Moss, S.E., Crumpton, M.J. (1988) Diversity in the lipocortin/calpactin family. Cell 55, 1–3

    Google Scholar 

  • Crumpton, M.J., Dedman, J.R. (1990) Protein terminology tangle. Nature 345, 212

    Google Scholar 

  • Cunninghame, M.E., Hall, J.L. (1986) The effect of calcium antagonists and inhibitors of secretory processes on auxin-induced elongation and fine structure of Pisum sativum stem segments. Protoplasma 133, 149–159

    Google Scholar 

  • Drust, D.S., Creutz, C.E. (1988) Aggregation of chromaffin granules by calpactin at micromolar levels of calcium. Nature 331, 88–91

    Google Scholar 

  • Felle, H. (1988) Auxin causes oscillations of cytosolic free calcium and pH in Zea mays coleoptiles. Planta 174, 495–499

    Google Scholar 

  • Gehring, C.A., Williams, D.A., Cody, S.H., Parish, R.W. (1990) Phototropism and geotropism in maize coleoptiles are spatially correlated with increases in cytosolic free calcium. Nature 345, 528–530

    Google Scholar 

  • Geisow, M.J., Walker, J.H. (1986) New proteins involved in cell regulation by Ca2+ and phospholipids. Trends Biochem. Sci. 11, 420–423

    Google Scholar 

  • Geisow, M.J., Walker, J.H., Boustead, C., Taylor, W. (1987) Annexins-new family of Ca2+ regulated phospholipid binding protein. Biosci. Rep. 7, 289–298

    Google Scholar 

  • Hong, K., Duzgunes, N., Papahadjopolous, D. (1981) Role of synexin in membrane fusion. J. Biol. Chem. 256, 3641–3644

    Google Scholar 

  • Jones, R.L., Robinson D.G. (1989) Protein secretion in plants. New Phytol. 111, 567–597

    Google Scholar 

  • Khanna, N.C., Helwig, E.D., Ikebuchi, N.W., Fitzpatrick, S., Bajwa, R., Waisman, D.M. (1990) Purification and characterisation of annexin proteins from bovine lung. Biochemistry 29, 4852–4862

    Google Scholar 

  • Kumar, B.V., Lakshmi, M.V., Atkinson, J.P. (1985) Fast and efficient method for detection and estimation of proteins. Biochem. Biophys. Res. Commun. 131, 883–891

    Google Scholar 

  • Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685

    PubMed  Google Scholar 

  • Miller, A.J., Sanders, D (1987) Depletion of cytosolic free Ca2+ induced by photosynthesis. Nature 326, 397–400

    Google Scholar 

  • Morris, M.R., Northcote, D.H. (1977) Influence of cations at the plasma membrane in controlling polysaccharide secretion from sycamore suspension cells. Biochem. J. 166, 603–618

    Google Scholar 

  • Nakata, T., Sobue, K., Hirokawa, N. (1990) Conformational change and localisation of calpactin I complex involved in exocytosis as revealed by quick-freeze, deep-etch electron microscopy and immunocytochemistry. J. Cell Biol. 110, 13–25

    Google Scholar 

  • Nobiling, R., Reiss, H.-D. (1987) Quantitative analysis of calcium gradients and activity in growing pollen tubes of Lilium longiflorum. Protoplasma 139, 20–24

    Google Scholar 

  • Picton, J.M., Steer, M.W. (1985) The effects of ruthenium red, lanthanum, fluorescein isothiocyanate and trifluoperazine on vesicle transport, vesicle fusion and tip extension in pollen tubes. Planta 163, 20–26

    Google Scholar 

  • Robertson, S., Potter, J.D. (1984) The regulation of free Ca2+ ion concentration by metal chelators. In: Methods in pharmacology, vol. 5: Myocardial biology, pp. 63–75, Schwartz, A., ed. Plenum Press New York London

    Google Scholar 

  • Sedmak, J.J., Grossberg, S.E. (1977) A rapid, sensitive and versatile assay for protein using Coomassie Brilliant Blue G250. Anal. Biochem. 79, 544–552

    Google Scholar 

  • Smallwood, M., Keen, J.N., Bowles, D.J. (1990) Purification and partial sequence analysis of plant annexins. Biochem. J. 279, 157–161

    Google Scholar 

  • Steer, M.W., Steer, J.M. (1990) Pollen tube tip growth. New Phytologist 111, 323–358

    Google Scholar 

  • Williams, D.A., Cody, S.H., Gehring, C.A., Parish, R.W., Harris, P.J. (1990) Confocal imaging of ionised calcium in living plant cells. Cell Calcium 11, 291–297

    Google Scholar 

  • Wilschut, J., Duzgunes, N., Fraley, R., Papahadjopoulos, D. (1980) Studies on the mechanism of membrane fusion: kinetics of calcium ion induced fusion of phosphatidylserine vesicles followed by a new assay for mixing of aqueous vesicle contents. Biochemistry 19, 6011–6021

    Google Scholar 

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Authors and Affiliations

  1. Department of Horticulture, School of Plant Sciences, University of Reading, Whiteknights, RG6 2AS, Reading

    H. D. Blackbourn & N. H. Battey

  2. Department of Biochemistry, University of Leeds, LS2 9JT, Leeds, UK

    J. H. Walker

Authors
  1. H. D. Blackbourn
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  2. J. H. Walker
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  3. N. H. Battey
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Additional information

This work was funded by the Agricultural and Food Research Council. Our thanks also to Professor P. Lowry and Dr R. Woods, Department of Biochemistry, University of Reading for facilities and advice for antiserum production, and C. Boustead, Department of Biochemistry, University of Leeds for advice on immunoblotting and phospholipid-binding assays.

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Blackbourn, H.D., Walker, J.H. & Battey, N.H. Calcium-dependent phospholipid-binding proteins in plants. Planta 184, 67–73 (1991). https://doi.org/10.1007/BF00208238

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