Theoretical and Applied Genetics

, Volume 71, Issue 1, pp 79–92 | Cite as

The structure and genetic control of a new class of disulphide-linked proteins in wheat endosperm

  • N. K. Singh
  • K. W. Shepherd


Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of unreduced total protein extracts from the endosperm of hexaploid wheat revealed three high molecular weight protein bands (triplet bands) in a zone of heavy background streaking. Electrophoretic examination of 135 hexaploid cultivars showed at least five different patterns of these triplet bands. Nine durum wheat cultivars showed a single band only. Analysis of nullisomic-tetrasomic and ditelocentric lines of ‘Chinese Spring’ wheat revealed that the slowest moving band (Tri-1) of the triplet was controlled by gene(s) on chromosome arm 1DS and the fastest moving band (Tri-3) by 1AS. The band with intermediate mobility (Tri-2) was found to be a hybrid aggregate of the subunits controlled by 1DS and 1AS. Using a non-reducing/reducing form of 2-dimensional (2-D) electrophoresis, these triplet bands were shown to be heterotetramers of four subunits designated D (M.W. 58,000), δ (22,000), A (52,000) and α (23,000) where Tri-1=DδDδ, Tri-2 = DδAα and Tri-3 = AαAα. With very low concentrations of 2-mercaptoethanol (ME), the tetramers dissociated into dimeric subunit pairs (Dδ, Aα), the monomers being observed with higher concentrations of ME. The structure of these subunit pairs resembles that of the subunit pairs in the globulin storage proteins of oats and some legumes. The 2-D method employed in this study was useful also for separating low molecular weight (LMW) subunits of glutenin from the monomeric gliadins which have similar electrophoretic mobility in 1-D separation. It was shown that at least four of these LMW glutenin subunits are controlled by genes on 1DS and 1AS and at least one subunit is controlled by gene(s) on 1BS. This electrophoretic separation method has proven useful in understanding the aggregation behaviour of the seed proteins of wheat.

Key words

Wheat Disulphide-linked proteins LMW glutenin subunits Genetic Control 2-D electrophoresis 


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  1. Beckwith AC, Wall JS, Jordon RW (1965) Reversible reduction and reoxidation of the disulphide bonds in wheat gliadin. Arch Biochem Biophys 112:16–24Google Scholar
  2. Beckwith AC, Nielsen HC, Huel JS, Huebner FR (1966) Isolation and characterization of high-molecular-weight protein from wheat gliadin. Cereal Chem 43:14–28Google Scholar
  3. Bietz JA, Wall JS (1972) Wheat gluten subunits: molecular weights determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Cereal Chem 49:416–430Google Scholar
  4. Bietz JA, Wall JS (1973) Isolation and characterization of gliadin-like subunits from glutenin. Cereal Chem 50: 537–547Google Scholar
  5. Bietz JA, Wall JS (1980) Identity of high molecular weight gliadin and ethanol-soluble glutenin subunits of wheat: relation to gluten structure. Cereal Chem 57:415–421Google Scholar
  6. Bietz JA, Shepherd KW, Wall JS (1975) Single-kernel analysis of glutenin: use in wheat genetics and breeding. Cereal Chem 52:513–532Google Scholar
  7. Bloksma AH (1975) Thiol and disulphide groups in dough rheology. Cereal Chem 52:170r-183rGoogle Scholar
  8. Booth MR, Ewart JAD (1969) Studies on four components of wheat gliadins. Biochim Biophys Acta 181:226–233Google Scholar
  9. Bottomley RC, Kearns HF, Schofield JD (1982) Characterization of wheat flour and gluten proteins using buffers containing sodium dodecyl sulphate. J Sci Food Agric 33: 481–491Google Scholar
  10. Brown JWS, Kemble RJ, Law CN, Flavell RB (1979) Control of endosperm proteins in Triticum aestivum (var ‘Chinese Spring’) and Aegilops umbellulata by homoeologous group 1 chromosomes. Genetics 93:189–200Google Scholar
  11. Casey R, March JF, Sanger E (1981) N-terminal amino acid sequence of α-subunits of legumin from Pisum sativum. Phytochemistry 20:161–163Google Scholar
  12. Charbonnier L (1974) Isolation and characterization of ω-gliadin fractions. Biochim Biophys Acta 359:142–151Google Scholar
  13. Crow MJA, Rothfus JA (1968) Chromatography of proteins from wheat gluten on polyacrylamide gel. Cereal Chem 45:413–420Google Scholar
  14. Danno G, Kanazawa K, Natake M (1974) Extraction of wheat flour proteins with sodium dodecyl sulphate and their molecular weight distribution. Agric Biol Chem 38: 1947–1953Google Scholar
  15. Galili G, Feldman M (1984) Mapping of glutenin and gliadin genes located on chromosome 1B of common wheat. Mol Gen Genet 193:293–298Google Scholar
  16. Gatehouse JA, Croy RRD, Boulter D (1980) Isoelectricfocusing properties and carbohydrate content of pea (Pisum sativum) legumin. Biochem J 185:497–503Google Scholar
  17. Huebner FR, Wall JS (1976) Fractionation and quantitative differences of glutenin from wheat varieties varying in baking quality. Cereal Chem 53:258–269Google Scholar
  18. Hynes RO, Destree A (1977) Extensive disulphide bonding at the mammalian cell surface. Proc Natl Acad Sci USA 74: 2855–2859Google Scholar
  19. Jackson EA, Holt LM, Payne PI (1983) Characterization of high molecular weight gliadin and low-molecular-weight glutenin subunits of wheat endosperm by two-dimensional electrophoresis and the chromosomal location of their controlling genes. Theor Appl Genet 66:29–37Google Scholar
  20. Jones IK, Philips JW, Hird FJR (1974) The estimation of Theologically important thiol and disulphide groups in dough. J Sci Food Agric 25:1–10Google Scholar
  21. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685Google Scholar
  22. Lásztity L (1984) Wheat proteins. In: The Chemistry of Cereal Proteins. CRC Press, Boca Raton Fla, pp 13–102Google Scholar
  23. Lawrence GJ, Payne PI (1983) Detection by gel electrophoresis of oligomers formed by the association of high-molecular-weight glutenin protein subunits of wheat endosperm. J Exp Bot 34:254–267Google Scholar
  24. Lawrence GJ, Shepherd KW (1980) Variation in glutenin protein subunits of wheat. Aust J Biol Sci 33:221–333Google Scholar
  25. Lawrence GJ, Shepherd KW (1981) Chromosomal location of genes controlling seed proteins in species related to wheat. Theor Appl Genet 59:25–31Google Scholar
  26. Matta N, Gatehouse JA, Boulter D (1981) The structure of legumin of Vicia faba L. — a reappraisal. J Exp Bot 32: 183–197Google Scholar
  27. Moreira MA, Hermodson MA, Larkins BA, Nielsen NC (1979) Partial characterization of the acidic and basic polypeptides of glycinin. J Biol Chem 254:9921–9926Google Scholar
  28. Nielsen HC, Babcock GE, Senti FR (1962) Molecular weight studies on glutenin before and after disulphide-bond splitting. Arch Biochem Biophys 96:252–258Google Scholar
  29. Nielsen HC, Beckwith AC, Wall JS (1968) Effect of disulphidebond cleavage on wheat gliadin fractions obtained by gel filtration. Cereal Chem 45:37–47Google Scholar
  30. Orth RA, Bushuk W (1974) Studies of glutenin. 6. Chromosomal location of genes coding for subunits of glutenin of common wheat. Cereal Chem 51:118–126Google Scholar
  31. Osborne TB (1907) Proteins of the wheat kernel. Carnegie Inst Washington Publ 84:1–119Google Scholar
  32. Payne PI, Corfield KG (1979) Subunit composition of wheat glutenin proteins isolated by gel filtration in a dissociating medium. Planta 145:83–88Google Scholar
  33. Payne PI, Law CN, Mudd EE (1980) Control by homoeologous group 1 chromosomes of the high-molecular-weight subunits of glutenin, a major protein of wheat endosperm. Theor Appl Genet 58:113–120Google Scholar
  34. Payne PI, Holt LM, Worland AJ, Law CN (1982) Structural and genetical studies on the high-molecular-weight subunits of wheat glutenin. 3. Telocentric mapping of the subunit genes on the long arms of homoeologous group 1 chromosomes. Theor Appl Genet 63:129–138Google Scholar
  35. Payne PI, Jackson EA, Holt LM, Law CN (1984) Genetic linkage between endosperm protein genes on each of the short arms of chromosomes 1A and 1B in wheat. Theor Appl Genet 67:235–243Google Scholar
  36. Pence JW, Olcott HS (1952) Effect of reducing agents on gluten proteins. Cereal Chem 29:292–298Google Scholar
  37. Peterson DM (1978) Subunit structure and composition of oat seed globulin. Plant Physiol 62:506–509Google Scholar
  38. Sears ER (1954) The aneuploids of common wheat. Mo Agric Expt Stn Res Bull 572:1–58Google Scholar
  39. Sears ER (1966) Nullisomic-tetrasomic combinations in hexaploid wheat. In: Riley R, Lewis KR (eds) Chromosome manipulations and plant genetics. Oliver and Boyd, Edinburgh, pp 29–45Google Scholar
  40. Shepherd KW (1968) Chromosomal control of endosperm proteins in wheat and rye. In: Finlay KW, Shepherd KW (eds) 3rd Int Wheat Genet Symp. Aust Acad Sci, Canberra, pp 86–96Google Scholar
  41. Singh NK, Shepherd KW (1984) A new approach to studying the variation and genetic control of disulphide-linked endosperm proteins in wheat and rye. In: Graveland A, Moonen JHE (eds) Proc 2nd Int Workshop Gluten Proteins. Wageningen, The Netherlands, pp 129–136Google Scholar
  42. Sommer A, Traut RR (1975) Identification by diagonal gel electrophoresis of nine neighboring protein pairs in the Escherichia coli 30 S ribosome crosslinked with methyl-4-mercaptobutyrimidate. J Mol Biol 97:471–481Google Scholar
  43. Walburg G, Larkins BA (1983) Oat seed globulin subunit characterization and demonstration of its synthesis as precursor. Plant Physiol 72:161–165Google Scholar
  44. Wang K, Richards FM (1974) An approach to nearest neighbor analysis of membrane proteins. Application to human erythrocyte membrane of a method employing cleavable cross-linkages. J Biol Chem 249:8005–8018Google Scholar
  45. Woychik JH, Huebner FR, Dimler RJ (1964) Reduction and starch-gel electrophoresis of wheat gliadin and glutenin. Arch Biochem Biophys 105:151–155Google Scholar
  46. Wrigley CW, Shepherd KW (1973) Electrofocusing of grain proteins from wheat genotypes. Ann NY Acad Sci 209: 154–162Google Scholar

Copyright information

© Springer-Verlag 1985

Authors and Affiliations

  • N. K. Singh
    • 1
  • K. W. Shepherd
    • 1
  1. 1.Department of AgronomyWaite Agricultural Research Institute, The University of AdelaideGlen OsmondAustralia

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