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The most prevalent protein in a heat-treated extract of pea (Pisum sativum) embryos is an LEA group I protein; its conformation is not affected by exposure to high temperature

Published online by Cambridge University Press:  19 September 2008

Pauline S. Russouw ,
Jill Farrant ,
Wolf Brandt  and
George G. Lindsey
Pauline S. Russouw
Affiliation:
Department of Biochemistry, University of Cape Town, Private Bag, Rondebosch 7700, South Africa
Jill Farrant
Affiliation:
Department of Botany, University of Cape Town, Private Bag, Rondebosch 7700, South Africa
Wolf Brandt
Affiliation:
Department of Biochemistry, University of Cape Town, Private Bag, Rondebosch 7700, South Africa
George G. Lindsey*
Affiliation:
Department of Biochemistry, University of Cape Town, Private Bag, Rondebosch 7700, South Africa
*
*Correspondence
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Abstract

The LEA-like protein previously isolated from a homogenate of pea (Pisum sativum L.) embryonic axes heated at 80°C for 10 min (Russouw et al., 1995) was purified without exposure to heat. Peptides produced by trypsin digestion were separated by HPLC and sequenced. The protein was identified as a member of the LEA group I family. The conformation of the protein was compared before and after heat treatment by antibody affinity, circular dichroism spectroscopy, fluorescence spectroscopy and 8-anilino-1-naphthalenesulfonic acid binding. No differences could be detected, demonstrating that the protein was not irreversibly denatured by exposure to high temperature.

Keywords

denaturationembryosheatLEA proteinpeas
Type
Biochemistry and Metabolism
Information
Seed Science Research , Volume 7 , Issue 2 , June 1997 , pp. 117 - 124
Copyright
Copyright © Cambridge University Press 1997

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References

Baker, J., Steele, C. and Dure, L. III (1988) Sequence and characterization of 6 LEA proteins and their genes from cotton. Plant Molecular Biology 11, 277–291.Google Scholar
Blackman, S.A., Wettlaufer, S.H., Obendorf, R.L. and Leopold, A.C. (1991) Maturation proteins associated with desiccation tolerance in soybean. Plant Physiology 96, 868–874.CrossRefGoogle ScholarPubMed
Blackman, S.A., Obendorf, R.L. and Leopold, A.C. (1995) Desiccation tolerance in developing soybean seeds: the role of stress proteins. Physiologia Plantarium 93, 630–638.CrossRefGoogle Scholar
Bradford, K.J. and Chandler, P.M. (1992) Expression of “dehydrin-like” proteins in embryos and seedlings of Zizania palustris and Oryza sativa during dehydration. Plant Physiology 99, 488–494.Google Scholar
Brand, L. and Witholt, B. (1967) Fluorescence measurements. Methods in Enzymology 11, 776–856.CrossRefGoogle Scholar
Brandt, W.F., Alk, H., Chauhan, M. and von Holt, C. (1984) A simple modification converts the spinning cup sequencer into a vapour phase sequencer. FEBS Letters 174, 228–232.Google Scholar
Close, T.J., Kort, A.A. and Chandler, P.M. (1989) A cDNA-based comparison of dehydration-induced proteins (dehydrins) in barley and corn. Plant Molecular Biology 13, 95–108.CrossRefGoogle ScholarPubMed
Close, T.J., Fenton, R.D. and Moonan, F. (1993) A view of plant dehydrins using antibodies specific to the carboxy terminal peptide. Plant Molecular Biology 23, 279–286.CrossRefGoogle Scholar
Dure, L. III, Crouch, M., Harada, J., Ho, T.H.D., Mundy, J., Quatrano, R., Thomas, T. and Sung, Z.R. (1989) Common amino acid sequence domains among the LEA proteins of higher plants. Plant Molecular Biology 12, 475–486.CrossRefGoogle ScholarPubMed
Espelund, M., Sæbøe-Larssen, S., Hughes, D.W., Galau, G.A., Larsen, F. and Jakobsen, K.S. (1992) Late embryogenesis-abundant genes encoding proteins with different numbers of hydrophilic repeats are regulated differentially by abscisic acid and osmotic stress. Plant Journal 2, 241–252.Google Scholar
Galau, G.A., Hughes, D.W. and Dure, L. III. (1986) Abscisic acid induction of cloned cotton late embryogenesis-abundant (LEA) mRNAs. Plant Molecular Biology 7, 157–170.Google Scholar
Galau, G.A., Wang, H.Y.C. and Hughes, D.W. (1992) Cotton Lea4 (D19) and LeaA2 (D132) Group 1 Lea genes encoding water stress-related proteins containing a 20-amino acid motif. Plant Physiology 99, 783–788.CrossRefGoogle ScholarPubMed
Guagliardi, A., Cerchia, L., Bartolucci, S. and Rossi, M. (1994) The chaperonin from the archeon Sulfolobus solfactaricus promotes correct refolding and prevents thermal denaturation in vitro. Protein Science 3, 1436–1443.CrossRefGoogle Scholar
Harlow, E. and Lane, D. (1988) Antibodies — a laboratory manual. Cold Spring Harbour Laboratory.Google Scholar
Hewick, R.M., Hunkapiller, M.W., Hood, L.E. and Dreyer, W.J. (1981) A gas-liquid solid phase peptide and protein sequencer. Journal of Biological Chemistry 256, 7990–7997.Google Scholar
Ingram, I. and Bartels, D. (1996) The molecular basis of dehydration tolerance in plants. Annual Review of Plant Physiology and Plant Molecular Biology 47, 377–403.Google Scholar
Jepson, S.G. and Close, T.J. (1995) Purification of a maize dehydrin protein expressed in Escherichia coli. Protein Expression and Purification 6, 632–636.Google Scholar
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of Bacteriophage T4. Nature 227, 680–685.CrossRefGoogle ScholarPubMed
Lindsey, G.G., Thompson, P., Pretorius, L., Purves, L.R. and von Holt, C. (1982) The effect of calcium ions on the stability of chicken erythrocyte histone octamers. FEBS Letters 149, 277–280.CrossRefGoogle Scholar
Lindsey, G.G., Thompson, P., Pretorius, L., Purves, L.R. and von Holt, C. (1983) Octamer reconstitution from acid extracted chicken erythrocyte histones. FEBS Letters 155, 301–305.CrossRefGoogle ScholarPubMed
Lottspiech, F. (1985) Microscale isocratic separation of the phenylthiohydantoin amino acid derivatives. Journal of Chromatography 326, 321–327.Google Scholar
McCubbin, W.D. and Kay, C.M. (1985) Hydrodynamic and optical properties of the wheat germ Em protein. Canadian Journal of Biochemistry and Cell Biology 63, 803–811.CrossRefGoogle Scholar
Mendoza, J.A., Lorimer, G.H. and Horowitz, P.M. (1992) Chaperonin cpn60 from Escherichia coli protects the mitochondrial enzyme rhodanese against heat inactivation and supports folding at elevated temperatures. Journal of Biological Chemistry 267, 17631–17634.Google Scholar
Russouw, P.S., Farrant, J.M., Brandt, W., Maeder, D. and Lindsey, G.G. (1995) Isolation and characterization of a heat-soluble protein from pea (Pisum sativum) embryos. Seed Science Research 5, 137–144.CrossRefGoogle Scholar
Stacy, R.A.P., Espelund, M., Sæbøe-Larsen, S., Hollung, K., Helliesen, E. and Jakobsen, K.S. (1995) Evolution of the Group I late embryogenesis abundant (Lea) genes: analysis of the Lea B19 gene family in barley. Plant Molecular Biology 28, 1039–1054.Google Scholar
Vertucci, C.W. and Farrant, J.M. (1995) Acquisition and loss of desiccation tolerance. pp 237–271 in Kigel, J. and Galili, G. (Eds). Seed development and germination. New York, Marcel Dekker Inc.Google Scholar