Abstract
As part of a study to identify potential targets of calcium action in plant cells, a 35-kDa, annexin-like protein was purified from pea (Pisum sativum L.) plumules by a method used to purify animal annexins. This protein, called p35, binds to a phosphatidylserine affinity column in a calcium-dependent manner and binds 45Ca2+ in a dot-blot assay. Preliminary sequence data confirm a relationship for p35 with the annexin family of proteins. Polyclonal antibodies have been raised which recognize p35 in Western and dot blots. Immunofluorescence and immunogold techniques were used to study the distribution and subcellular localization of p35 in pea plumules and roots. The highest levels of immunostain were found in young developing vascular cells producing wall thickenings and in peripheral root-cap cells releasing slime. This localization in cells which are actively involved in secretion is of interest because one function suggested for the animal annexins is involvement in the mediation of exocytosis.
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Abbreviations
- SDS-PAGE:
-
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
References
Bennet, R.J., Breen, C.M., Bandu, V.H. (1990) A role for calcium in the cellular differentiation of root cap cells: A re-examination of root growth control mechanisms. Environ. Exp. Bot. 30, 515–523
Blackbourn, H.D., Walker, J.H., Battey, N.H. (1991) Calcium dependent phospholipid-binding proteins in plants. Planta 184, 67–73
Brutlag, D.L., Dautrico, J.P., Maulik, S., Relph, J. (1990) Improved sensitivity of biological sequence data base searches. Comp. Appl. Biol. Sci. 6, 237–245
Burgoyne, R.D., Giesow, M.J. (1989) The annexin family of calcium-binding proteins. Cell Calcium 10, 1–10
Clark, G.B., Dauwalder, M., Roux, S.J. (1989) Partial purification and characterization of a calcium-phospholipid binding protein in peas. (Abstr.) J. Cell Biol. 4, 304a
Clark, G.B., Dauwalder, M., Roux, S.J. (1990) Purification, Biochemical characterization, and localization of a calciumphospholipid binding protein in peas. (Abstr.) J. Cell Biol. 5, 356a
Cleveland, D.W., Fischer, S.G., Kirschner, M.W., Laemmli, U.K. (1977) Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J. Biol. Chem. 252, 1102–1106
Crompton, M.R., Moss, S.E., Crumpton, M.J. (1988) Diversity in the lipocortin/calpactin family Cell 55, 1–3
Dauwalder, M., Whaley, W.G. (1974) Patterns of incorporation of [3H]galactose by cells of Zea mays root tips. J. Cell Sci. 14, 11–27
Dauwalder, M., Whaley, W.G., Kephart, J.E. (1969) Phosphatases and differentiation of the Golgi apparatus. J. Cell Sci. 4, 455–498
Dauwalder, M., Roux, S.J., Hardison L. (1986) Distribution of calmodulin in pea seedlings: Immunocytochemical localization in plumules and root apices. Planta 168, 461–470
Dayhoff, M.O., Eck, R.V., Park, C.M. (1972) A model of evolutionary change in proteins. In: Atlas of protein sequence and structure 1972. vol. 5, pp. 89–99, Dayhoff, M.O., ed. National Biomedical Research Foundation Silver Spring, Md., USA
Dedman, J. (1986) Mediation of intracellular calcium: Variances on a common theme. Cell Calcium 7, 297–307
Friedmann, M., Poovaiah, B.W. (1991) Calcium and protein phos phorylation in the transduction of gravity signal in corn roots. Plant Cell Physiol. 32, 299–302
Gerke, V., Koch, W. (1990) The cDNA sequence of chicken annexin II. Nucleic Acids Res. 18, 4246
Haigler, H.T., Christmas, P. (1990) Annexin I is secreted by the human prostate. Biochem. Soc. Trans. 18, 1104–1106
Kyhse-Anderson, J. (1984) Electroblotting of multiple gels: a simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide to nitrocellulose. J. Biophys. Biochem. Methods 10, 203–209
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685
Lillie, S.H., Brown, S.S. (1987) Artifactual immunofluorescent la belling in yeast, demonstrated by affinity purification of anti body. Yeast 3, 63–70
Martin, L.N. (1982) Separation of guinea pig IgG subclasses by affinity chromatography on protein A-sepharose. J. Immunol. Methods 52, 205–212
Maruyama, K., Mikawa, T., Ebashi, S. (1984) Detection of calcium binding proteins by 45Ca autoradiography on nitrocellulose membrane after sodium dodecyl sulfate gel electrophoresis. J. Biochem. 95, 511–519
Moore, R., Fondren, W.M. (1988) A gradient of endogenous calcium forms in mucilage of graviresponding roots of Zea mays. Ann. Bot. 61, 113–116
Oakley, B.R. (1980) A simplified ultrasensitive silver stain for de tecting proteins in polyacrylamide gels. Anal. Chem. 105, 361–363
O'Farrell, P.H. (1975) High resolution two-dimensional electrophoresis of proteins. J. Biol. Chem. 250, 4007–4021
Poovaiah, B.W., McFadden, J.J., Reddy, A.S.N. (1987) The role of calcium-ions in gravity signal perception and transduction. Physiol. Plant. 71, 401–407
Roberts, A.W., Haigler, C.H. (1989) Tracheary-element differentiation in suspension-cultured cells of Zinnia requires uptake of extracellular Ca2+. Planta 180, 502–509
Roberts, D.M., Lukas, T.J., Watterson, D.M. (1986) Structure, function, and mechanism of action of calmodulin. CRC Crit. Rev. Plant Sci. 4, 311–339
Roux, S.J., Serlin, B.S. (1987) Cellular mechanisms controlling light-stimulated gravitropism: role of calcium. CRC Crit. Rev. Plant Sci. 5, 205–236
Smallwood, M., Keen, J.N., Bowles, D.J. (1990) Purification and partial sequence analysis of plant annexins. Biochem J. 270, 157–161
Steer, M.W. (1988) The role of calcium in exocytosis and endocy tosis in plant cells. Physiol. Plant. 72, 213–220
Uchida, T., Filburn, C.R. (1984) Affinity chromatography of protein kinase C-phorbol ester receptor on polyacrylamide immobilized phosphatidylserine. J. Biol. Chem. 259, 12311–12314
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The authors thank Dennis Brown (Cell Research Institute) for use of this research facilities and Zainab Ilahi and Collin Thomas for valuable technical assistance. Portions of this work were presented at the 1989 and 1990 meetings of the American Society for Cell Biology (Clark et al. 1989, 1990). This work was in part supported by a National Institute of Health Training Grant 1-T32-HD07296-01A3 and by a National Aeronautics and Space Administration grant NAGW 1519.
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Clark, G.B., Dauwalder, M. & Roux, S.J. Purification and immunolocalization of an annexin-like protein in pea seedlings. Planta 187, 1–9 (1992). https://doi.org/10.1007/BF00201617
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DOI: https://doi.org/10.1007/BF00201617