Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Identification and isolation of methionine-cysteine rich proteins in soybean seed

Abstract

We recently developed a method to identify methionine-containing proteins and quantitate their methionine contents [4]. We applied this method to soybeans and identified relatively methionine-rich proteins (MRP) among the albumins. By acidic methanol extraction of the albumins, we obtained a group of low molecular weight methionine-cysteine rich proteins (MCRP) that analyzed 4.0% methionine and 8.8% cysteine. MCRP made up 1–2% of the total protein in soybeans. Reversed-phase HPLC purification of MCRP yielded a protein peak that exhibited a single major band on denaturing polyacrylamide gel electrophoresis, had a molecular weight of 16kD and contained 6.2% methionine and 18.8% cysteine. We are cloning the gene for this protein. Increasing its level through genetic engineering could increase the methionine-cysteine content of soybeans.

This is a preview of subscription content, log in to check access.

References

  1. 1.

    Altenbach SB, Pearson KW, Leung FW, Sun SSM (1987) Cloning and sequence analysis of a cDNA encoding a Brazil nut protein exceptionally rich in methionine. Plant Mol Biol 8: 239–250

  2. 2.

    Apostolatos G (1984) Isolation and characterization of a methionine-rich protein fraction from edible dry bean (Phaseolus vulgaris L.). Plant Sci Letters 33: 39–46

  3. 3.

    Birk Y (1961) Purification and some properties of a highly active inhibitor of trypsin and chymotrypsin from soybean. Biochim Biophys Acta 54: 378–381

  4. 4.

    de Lumen BO, Kho CJ (1987) Identification of their methionine contents. J Agric Food Chem 35: 688–691

  5. 5.

    FAO/WHO Tech. Rep. Ser. No. 522 (1973) p. 63

  6. 6.

    Foard DE, Gutay PA, Ladin B, Beachy RN, Larkins BA (1982) In vitro synthesis of the Bowman-Birk and related soybean protease inhibitors. Plant Mol Biol 1: 227–243

  7. 7.

    Frattali V (1969) Soybean inhibitors III properties of a low molecular weight soybean proteinase inhibitor. J. Bio Chem 244: 274–280

  8. 8.

    Higgins TJV (1986) Personal communication

  9. 9.

    Higgins TJV, Chandler PM, Randall PJ, Spencer D, Beach LR, Blagrove RJ, Kort AA, Inglis AS (1986) Gene structure, protein structure and regulation of a sulfur-rich protein in pea seeds. J Biol Chem 261: 11124–11130

  10. 10.

    Hirs CHW (1967) Determination of cysteine as cysteic acid. In: Hirs CHW (ed.) Methods in Enzymology. New York: Academic Press, pp. 59–62

  11. 11.

    Holowach LP, Thompson JF, Madison JT (1984) Effects of exogenous methionine on storage protein composition of soybean cotyledons cultured in vitro. Plant Physiol 74: 576–579

  12. 12.

    Hwang DLR, Davis Lin KT, Yang WK, Foard DE (1977) Purification, partial characterization, and immunological relationships of multiple low molecular weight protease inhibitors of soybean. Biochim Biophys Acta 495: 369–382

  13. 13.

    Jaynes J, Yang M, Espinoza M, Dodds J (1986) Plant protein improvement by genetic engineering: use of synthetic genes. Trends in Biotech 4: 314–320

  14. 14.

    Kakade ML, Rackis JJ, McGhee JE, Puski G (1974) Determination of trypsin inhibitor activity of soy products: a collaborative analysis of an improved procedure. Cereal Chem 51: 376–379

  15. 15.

    Koshiyama I (1968) Chemical and physical properties of a 7S protein in soybean globulin. Cereal Chem 45: 394–404

  16. 16.

    Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685

  17. 17.

    Millar DBS, Willick GE, Steiner RF, Frattali V (1969) Soybean Inhibitors IV. The reversible self-association of a soybean proteinase inhibitor. J Biol Chem 244: 281–284

  18. 18.

    Murray DR (1979) A storage role for the albumins in pea cotyledons. Plant, Cell and Environment 2: 221–226

  19. 19.

    Nielsen NC (1984) The chemistry of legume storage proteins. Phil Trans R Soc Lond B304: 287–296

  20. 20.

    Odani S, Ikenaka T (1977) Studies on soybean trypsin inhibitors X. Isolation and partial characterization of four soybean double-headed proteinase inhibitors. J Biochem (Tokyo) 82: 1513–1522

  21. 21.

    Odani S, Ikenaka T (1978) Studies on soybean trypsin inhibitors XIII. Preparation and characterization of active fragments from Bowman-Birk proteinase inhibitor. J Biochem (Tokyo) 83: 747–753

  22. 22.

    Rackis J (1961) Amino acids in soybean hulls and oilmeal fractions. J Agric Food Chem 9: 409–412

  23. 23.

    Shvarts VS, Vaintraub IA (1967) isolation of the 11S component of soya bean protein and determination of its amino acid composition by an automatic chromato-polarographic method. Biokhimiya 32: 162–168; Biochemistry (USSR) (English Translation) 32: 135–140

  24. 24.

    Staswick PE, Hermodson MA, Nielsen NC (1984) The amino acid sequence of the A2B1a subunit of glycinin. J Biol Chem 259: 13424–13430

  25. 25.

    Sun SSM, Leung FM, Tomic JC (1987) Brazil nut (Bertholletia excelsa H.B.K.) proteins: Fractionation, composition and identification of a sulfur-rich protein. J Agric Food Chem 35: 232–235

  26. 26.

    Youle RJ, Huang AHC (1981) Occurrence of low molecular weight high-cysteine containing albumin storage proteins in oilseeds of diverse species. Amer J Bot 68: 44–48

Download references

Author information

Correspondence to Benito O. de Lumen.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Kho, C., de Lumen, B.O. Identification and isolation of methionine-cysteine rich proteins in soybean seed. Plant Food Hum Nutr 38, 287–296 (1988). https://doi.org/10.1007/BF01091726

Download citation

Key words

  • soybean
  • methionine-cysteine rich protein