Abstract
Serological studies and comparison of N-terminal amino acid sequences with the amino acid sequence deduced from a cDNA clone have been used to establish the sequence relationships between the subunits of the pea seed storage protein, vicilin. Subunits smaller than Mr∼50 000 (i.e., Mr 34 000, 30 000, 25 000, 18 000, 14 000, 13 000 and 12 000) show extensive homology with molecules within Mr∼50 000 group. Both the sequencing and serological data confirm earlier evidence from studies on vicilin synthesisin vivo andin vitro which indicated that the vicilin subunits smaller than Mr∼50 000 arose by endoproteolytic cleavage of parent molecules within the Mr∼50 000 group. Cleavage in different Mr 50 000 parent molecules containing either one or both of two susceptible processing sites accounts for the formation of all the vicilin subunits smaller than Mr∼50 000, with the possible exception of the Mr34 000 polypeptide. The position of these sites in the putative parents were defined by reference to a complete amino acid sequence deduced from the sequence of DNA complementary to mRNA for one member of the Mr∼50 000 group.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Badenoch-Jones J, Spencer D, Higgins TGV, Millerd A: The role of glycosylation in storage-protein synthesis in developing pea seeds. Planta 153:201–209, 1981.
Bittner M, Kupferer P, Morris CE: Electrophoretic transfer of proteins and nucleic acids from slab gels to diazobenzyloxymethyl cellulose or nitrocellulose sheets. Anal Biochem 102:359–471, 1980.
Blagrove RJ, Gillespie JM: Isolation, purification and characterization of the seed globulins of Lupinus angustifolius. Aust J Plant Physiol 2:13–27, 1975.
Chandler PM, Higgins TJV, Randall PJ, Spencer D: Regulation of legumin levels in developing pea seeds under conditions of sulfur deficiency: rates on legumin synthesis and levels of legumin mRNA. Plant Physiol 71:47–54, 1983.
Chrispeels MJ, Higgins TJV, Spencer D: Assembly of storage protein oligomers in the endoplasmic reticulum and processing of the polypeptides in the protein bodies of developing pea cotyledons. J Cell Biol 93:306–313, 1982.
Croy PRD, Gatehouse JA, Evans DM, Boulter D: Characterization of the storage protein subunits synthesized in vitro by polyribosomes and RNA from developing pea (Pisum sativum L.) 1. Legumin. Planta 148:49–56, 1980.
Davey RA, Higgins TJV, Spencer D: Homologies between two small subunits of vicilin from Pisum sativum. Biochem Internat 3:595–602, 1981.
Derbyshire E, Wright DJ, Boulter D: Legumin and vicilin, storage proteins of legume seeds. Phytochemistry 15:3–24, 1976.
Gatehouse JA, Croy RRD, Morton H, Tyler M, Boulter D: Characterisation and subunit structures of the vicilin storage proteins of pea (Pisum sativum L.). Eur J Biochem 118:627–633, 1981.
Gatehouse JA, Lycett GW, Croy RRD, Boulter D: The post-translational proteolysis of the subunits of vicilin from pea (Pisum sativum L.). Biochem J 207:629–632, 1982.
Higgins TJV, Chandler PM, Zurawski G, Button SC, Spencer D: The biosynthesis and primary structure of pea seed lectin. J Biol Chem 258:9544–9549, 1983.
Higgins TJV, Spencer D: Precursor forms of pea vicilin subunits. Modifications by microsomal membranes during cell-free translation. Plant Physiol 67:205–211, 1981.
Hirano H, Gatehouse JA, Boulter D: The complete amino acid sequence of a subunit of the vicilin seed storage protein of pea (Pisum sativum L.). FEBS Lett 145:99–102, 1982.
Inglis AS: Modified extraction procedures in spinning-cup sequenators. In: Hirs CWH, Timasheff SN (eds) Methods in Enzymology, 91. Academic Press, New York, 1983. pp 443–450.
Maxam AM, Gilbert W: Sequencing end-labelled DNA with base-specific chemical cleavages. In: Grossman L, Moldave K (eds) Methods in Enzymology, 65. Academic Press, New York, 1980, pp 499–560.
Millerd A, Spencer D: Changes in RNA-synthesizing activity and template activity in nuclei from cotyledons of developing pea seeds. Aust J Plant Physiol 1:331–341, 1974.
Spencer D, Higgins TJV: The biosynthesis of legumin in maturing pea seeds. Biochem Internat 1:502–509, 1980.
Spencer D, Higgins TJV, Button SC, Davey RA: Pulse-labelling studies on protein synthesis in developing pea seeds and evidence of a precursor form of legumin small subunit. Plant Physiol 66:510–515, 1980.
Struck DK, Lennarz WJ, Brew K: Primary structural requirements for the enzymatic formation of the N-glycosidic bond in glycoproteins. J Biol Chem 253:5786–5794, 1978.
Sun SM, McLeester RC, Bliss FA, Hall TC: Reversible and irreversible dissociation of globulins from Phaseolus vulgaris seed. J Biol Chem 249:2118–2120, 1974.
Thanh VH, Shibasaki K: Major proteins of soybean seeds. Subunit structure of β-conglycinin. J Agric Food Chem 26:692–695, 1978.
Thomson JA, Schroeder HE, Dudman WE: Cotyledonary storage proteins in Pisum sativum. I. Molecular heterogeneity. Aust J Plant Physiol 5:263–279, 1978.
Thomson JA, Schroeder HE, Tassie AM: Cotyledonary storage proteins in Pisum sativum. V. Further studies on molecular heterogeneity in the vicilin series of holoproteins. Aust J Plant Physiol 7:271–282, 1980.
Towbin H, Staehlin T, Gordon J: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354, 1979.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Spencer, D., Chandler, P.M., Higgins, T.J.V. et al. Sequence interrelationships of the subunits of vicilin from pea seeds. Plant Mol Biol 2, 259–267 (1983). https://doi.org/10.1007/BF01578644
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01578644