Summary
TheRicinus communis (castor bean) 2S albumin is a heterodimer of glutamine-rich, disulphidelinked 4 and 7 kDa polypeptides. A cDNA library was constructed using mRNA from maturing castor bean endosperm as template. Clones containing sequences complementary to albumin mRNA were isolated by hybridization using as a probe a mixture of synthetic oligonucleotides representing sequences predicted for a peptide present in the 2 S albumin large subunit. The nucleotide sequence contained an open reading frame encoding a preproprotein of 258 amino acid residues. The preproprotein included both polypeptides of the previously sequenced 2S albumin. In addition, this precursor included two further glutamine-rich sequences which, in terms of their size and conserved cysteine residues typically found in seed proteins of the 2S albumin superfamily, possibly represent the small and large polypeptide subunits of a second heterodimeric storage protein. A post-translational processing scheme is proposed which would result in a single preproprotein generating two distinct heterodimeric 2S albumins. The generation of a second heterodimer seems likely since polypeptide candidates for its small and large subunits were found in theRicinus 2S albumin fraction, and N-terminal protein sequencing confirmed the existence of the putative small subunit.
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Allen RD, Cohen EA, Vonder Haar RA, Adams CA, Ma DP, Nessler CL, Thomas TL (1987) Sequence and expression of a gene encoding an albumin storage protein in sunflower. Mol Gen Genet 210:211–218
Altenbach SB, Pearson KW, Leung FW, Sun SSM (1987) Cloning and sequence analysis of a cDNA encoding a Brazil nut protein extremely rich in methionine. Plant Mol Biol 8:239–250
Anderson CW, Straus JW, Dudock BS (1983) Preparation of a cell-free protein-synthesizing system from wheat germ. Methods Enzymol 101:635–644
Aviv H, Leder P (1972) Purification of biologically active globin messenger RNA by chromatography on oligothymidilic acidcellulose. Proc Natl Acad Sci USA 69:1408–1412
Boulter D (1984) Cloning of pea storage protein genes. Philos Trans R Soc Lond [Biol] 304:323–332
Bowles DJ, Marcus SE, Pappin DJC, Findlay JBC, Eliopolous E, Maycox PR, Burgess J (1986) Posttranslational processing of concanavalin A precursors in jackbean cotyledons. J Cell Biol 102:1284–1297
Butterworth AG, Lord JM (1983)Ricin andRicinius communis agglutinin subunits are all derived from single sized precursor polypeptides. Eur J Biochem 137:57–65
Campos FAP, Richardson M (1983) The complete amino acid sequence of the bifunctional α-amylase/trypsin inhibitor from seeds of ragi (Indian finger millet,Elusine coracana). FEBS Lett 152:300–304
Chan RL, Keller M, Canaday J, Weil J-H, Imbault P (1990) Eight small subunits ofEuglena ribulose-1,5-bisphosphate carboxylase/oxygenase are translated from a large mRNA as a polyprotein. EMBO J 9:333–338
Crouch ML, Tenbarge KM, Simon AE, Ferl R (1983) cDNA clones forBrassica napus seed storage proteins: evidence from nucleotide sequence analysis that both subunits of napin are cleaved from a precursor polypeptide. J Mol Appl Genet 2:273–283
Dickinson CD, Evans RP, Nielson NC (1988) RY repeats are conserved in the 5' flanking regions of legume seed protein genes. Nucleic Acids Res 16:371
Ericson ML, Rodin J, Lenman M, Glimelius K, Josefsson L-G, Rask L (1986) Structure of the rape seed 1.7S storage protein, napin, and its precursor. J Biol Chem 261:14576–14581
Findlay JBC, Pappin DJC, Keen JN (1989) Automated solid-phase microsequencing. In: Findlay JBC, Geisow M (eds) Protein sequencing — A practical approach. IRL Press, Oxford, pp 69–84
Forde J, Forde BG, Fry RP, Kreis M, Shewry PR, Miflin BJ (1983) Identification of barley and wheat cDNA clones related to the high Mr polypeptides of wheat gluten. FEBS Lett 162:360–366
Halpin C, Conder MJ, Lord JM (1989) Different routes for integral protein insertion intoRicinus communis protein body and glyoxysome membranes. Planta 179:331–339
Higgins TJV (1984) Synthesis and regulation of major proteins in seeds. Annu Rev Plant Physiol 35:191–221
Higgins TJV, Chandler PM, Zurawski G, Button S, Spencer D (1983) The biosynthesis and primary structure of pea seed lectin. J Biol Chem 258:9544–9549
Holmes DS, Quigley M (1981) A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114:193–197
Houlne G, Schartz R (1988) Characterization of cDNA sequences for LHCI apoproteins inEuglena gracilis: the mRNA encodes a large precursor containing several consecutive divergent polypeptides. Mol Gen Genet 213:479–486
Irwin SD, Lord JM (1990) Nucleotide sequence of aRicinus communis 2S albumin precursor gene. Nucleic Acids Res
Iyengar RB, Smits P, van der Ouderaa F, van der Well H, van Browershaven J, Ravestein P, Richters G, van Wassenaar PD (1979) The complete amino acid sequence of the sweet protein thaumatin 1. Eur J Biochem 96:193–204
Josefsson L-G, Lenman M, Ericson ML, Rask L (1987) Structure of a gene encoding the 1.7S storage protein, napin, fromBrassica napus. J Biol Chem 262:12196–12201
Krebbers E, Herdies L, de Clerq A, Seurinck J, Leemans J, van Damme J, Segura M, Gheysen G, van Montagu M, Vanderkerckhove J (1988) Determination of the processing sites of anArabidopsis 2S albumin and characterization of the complete gene family. Plant Physiol 87:859–866
Kreis M, Shewry PR, Forde BG, Forde J, Miflin BJ (1985) Structure and evolution of seed storage proteins and their genes, with particular reference to wheat, barley and rye. Oxford Surv Plant Mol Cell Biol 2:253–317
Lamb FI, Roberts LM, Lord JM (1985) Nucleotide sequence of cloned cDNA coding for preproricin. Eur J Biochem 148:265–270
Laursen RA (1971) Solid phase Edman degradation. An automatic peptide sequencer. Eur J Biochem 20:89–102
Lehrach H, Frischauf AM, Hanahan D, Wozney J, Fuller F, Boedtker H (1979) Construction and characterization of prα1 collagen complementary deoxyribonucleic acid clones. Biochemistry 18:3146–3152
Lord JM (1985) Synthesis and intracellular transport of lectin and storage protein precursors in endosperm from castor bean. Eur J Biochem 146:403–409
Lord JM, Robinson C (1986) The role of proteolytic enzymes in the posttranslational modification of proteins. In: Dalling MJ (ed) Plant proteolytic enzymes. CRC Press, Boca Raton, Fla, pp 69–80
Maeda K, Hase T, Matsubara H (1983) Complete amino acid sequence of an α-amylase inhibitor in wheat kernel. Biochim Biophys Acta 743:52–57
Mahoney WC, Hermodson MA, Jones B, Powers DD, Corfman RS, Reech GR (1984) Amino acid sequence and secondary structure analysis of the corn inhibitor of trypsin and activated Hageman factor. J Biol Chem 259:8412–8416
McGurl B (1987) Purification and characterization of putative protein allergens from the seeds of the castor oil plant,Ricinus communis. PhD Thesis, University of Warwick, UK
Melton DA, Krieg PA, Rebagliati MR, Maniatis T, Zinn K, Green MR (1984) Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res 12:7035–7056
Odani S, Koide T, Ono T (1983) The complete amino acid sequence of barley trypsin inhibitor. J Biol Chem 258:7998–8003
Roberts LM, Lord JM (1981 a) Protein biosynthetic capacity in the endosperm tissue of ripening castor bean seeds. Planta 152:420–427
Roberts LM, Lord JM (1981 b) The synthesis ofRicinius communis agglutinin. Cotranslational and posttranslational modification of agglutinin polypeptides. Eur J Biochem 119:31–41
Roberts LM, Lamb FI, Pappin DJC, Lord JM (1985) The primary sequence ofRicinus communis agglutinin. Comparison with rice. J Biol Chem 260:15682–15686
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Scofield SR, Crouch ML (1987) Nucleotide sequence of a member of the napin storage protein family fromBrassica napus. J Biol Chem 262:12202–12208
Sharief FS, Li SS-L (1982) Amino acid sequence of the small and large subunits of seed storage proteins fromRicinius communis. J Biol Chem 257:14753–14759
Tan CGL, Stevens FC (1971) Amino acid sequence of lima bean protease inhibitor component IV. 2. Isolation, sequence determination of the chymotryptic peptides and the complete amino acid sequence. Eur J Biochem 18:515–523
Von Heijne G (1984) How signal sequences maintain cleavage specificity. J Mol Biol 173:243–251
Youle RJ, Huang AHC (1978) Albumin storage proteins in the protein bodies of castor bean. Plant Physiol 61:13–16
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Irwin, S.D., Keen, J.N., Findlay, J.B.C. et al. Theinus communis2S albumin precursor: A single preproprotein may be processed into two different heterodimeric storage proteins. Mol Gen Genet 222, 400–408 (1990). https://doi.org/10.1007/BF00633846
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DOI: https://doi.org/10.1007/BF00633846