Skip to main content

Advertisement

Log in

A proteomic approach to the identification and characterisation of protein composition in wheat germ

  • Original Paper
  • Published:
Functional & Integrative Genomics Aims and scope Submit manuscript

Abstract

Proteome analyses were carried out on commercial wheat germ of mature grain from the biscuit-making wheat cultivar, Rosella. Wheat germ protein extracts were fractionated by two-dimensional gel electrophoresis across two different immobilised pH gradients: pH 4.0–7.0 and 6.0–9.0. A total of 612 individual protein spots were excised from the gels and characterised by peptide mass fingerprinting. From these analyses, 347 individual proteins were identified from protein sequence database interrogation, and 301 different types of protein were catalogued according to protein function. The remaining 265 protein spots gave poor or no matches to proteins in the databases and were not identified in this study. Six different classes of enzymes were identified in the germ, many of them having roles in the mobilisation of energy reserves for germination. Abundantly expressed enzyme classes include the oxidoreductases, transferases and hydrolases. A comparison was also made between the major protein classes expressed in the germ and protein classes expressed in the endosperm from previous proteomic work. This study contributes significantly to our knowledge of protein expression and heterogeneity in the germ of wheat grain and forms the basis for future studies in regard to the characterisation of proteins during the initial stages of germination.

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

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2

Similar content being viewed by others

Abbreviations

AcN:

Acetonitrile

2-DE:

two-dimensional gel electrophoresis

IEF:

isoelectric focussing

IPG:

immobilised pH gradient

MALDI-TOF:

matrix-assisted laser desorption/ionisation-time of flight

PMF:

peptide mass fingerprinting

SDS-PAGE:

sodium dodecyl sulphate–polyacrylamide gel electrophoresis

TBP:

tributyl phosphine

TCA:

trichloroacetic acid

TFA:

trifluoroacetic acid

References

  • Amiour N, Merlino M, Leroy P, Branlard G (2002) Proteomic analysis of amphiphilic proteins of hexaploid wheat kernels. Proteomics 2:632–641

    Article  PubMed  CAS  Google Scholar 

  • Bevan M, Bancroft I et al (1998) Analysis of 1.9 Mb of contiguous sequence from chromosome 4 of Arabidopsis thaliana. Nature 391:485–488

    Article  PubMed  CAS  Google Scholar 

  • Branlard G, Amiour N, Igrejas G, Gaborit T, Herbette S, Dardevet M, Marion D (2003) Diversity of puroindolines as revealed by two-dimensional electrophoresis. Proteomics 3:168–174

    Article  PubMed  CAS  Google Scholar 

  • Clarke BC, Hobbs M, Skylas D, Appels R (2000) Genes active in developing wheat endosperm. Funct Integr Genomics 1:44–55

    Article  PubMed  CAS  Google Scholar 

  • Cornish GB, Skylas DJ, Siriamornpun S, Bekes F, Wrigley CW, Wootton M (2001) Grain proteins as markers of genetic traits in wheat. Aust J Agric Res 52:1161–1171

    Article  CAS  Google Scholar 

  • Cozzolino R, DiGiorgi S, Fisichella S, Garozzo D, Lafiandra D, Palermo A (2001) Proteomics of gluten: mapping of subunit 1 A×2* in Cheyenne cultivar by matrix-assisted laser desorption/ionisation. Rapid Commun Mass Spectrom 15:1129–1135

    Article  PubMed  CAS  Google Scholar 

  • Granier F (1988) Extraction of plant proteins for two-dimensional electrophoresis. Electrophoresis 9:712–718

    Article  PubMed  CAS  Google Scholar 

  • Gorg A, Obermaier C, Boguth G, Harder A, Scheibe B, Wildgruber R, Weiss W (2000) The current state of two-dimensional electrophoresis with immobilised pH gradients. Electrophoresis 21:1037–1053

    Article  PubMed  CAS  Google Scholar 

  • Fu BX, Sapirstein HD (1996) Procedure for isolating monomeric and polymeric glutenin of wheat. Cereal Chem 73:143–152

    CAS  Google Scholar 

  • Khatkar BS, Schofield JD (1997) Molecular and physicochemical basis of breadmaking—properties of wheat gluten proteins: a critical appraisal. J Food Sci Technol 34:85–102

    CAS  Google Scholar 

  • Majoul T, Bancel E, Triboi E, Ben Hamida J, Branlard G (2004) Proteomic analysis of the effect of heat stress on hexaploid wheat grain: characterization of heat-responsive proteins from non-prolamins fraction. Proteomics 4:505–513

    Article  PubMed  CAS  Google Scholar 

  • Marcus A, Efron D, Weeks DP (1974) The wheat embryo cell-free system. Methods Enzymol 30:749–754

    Article  PubMed  CAS  Google Scholar 

  • Mares DJ, Mrva K (2001) Mapping quantitative trait loci associated with variation in grain dormancy in Australian wheat. Aust J Agric Res 52:1257–1265

    Article  CAS  Google Scholar 

  • Mikami S, Kishimoto T, Hori H, Mitsui T (2002) Technical improvement to 2D-PAGE of rice organelle membrane proteins. Biosci Biotechnol Biochem 66:1170–1173

    Article  PubMed  CAS  Google Scholar 

  • Neuhoff V, Arold N, Taube D, Ehrhardt W (1988) Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie brilliant blue G-250 and R-250. Electrophoresis 9(6):255–262

    Article  PubMed  CAS  Google Scholar 

  • Nouwens AS, Cordwell SJ, Larsen MR, Molloy MP, Gillings M, Willcox MDP, Walsh BJ (2000) Complementing genomics with proteomics: the membrane subproteome of Pseudomonas aeruginosa. Electrophoresis 21:3797–3809

    Article  PubMed  CAS  Google Scholar 

  • Shimoni Y, Zhu Xz, Levanony H, Segal G, Galili G (1995) Purification, characterization, and intracellular localization of glycosylated protein disulfide isomerase from wheat grains plant. Physiology 108(1):327–334

    CAS  Google Scholar 

  • Skylas DJ, Mackintosh JA, Cordwell SJ, Basseal DJ, Walsh BJ, Harry J, Blumenthal C, Copeland L, Wrigley CW, Rathmell W (2000) Proteome approach to the characterisation of protein composition in the developing and mature wheat-grain endosperm. J Cereal Sci 32:169–188

    Article  CAS  Google Scholar 

  • Skylas DJ, Van Dyk D, Wrigley CW (2005) Proteomics of wheat grain. J Cereal Sci 41:165–179

    Article  CAS  Google Scholar 

  • Tatham AS, Shewry PR, Belton PS (1990) Structural studies of cereal prolamins, including wheat gluten. In: Pomeranz Y (ed) Advances in cereal science and technology. American Association of Cereal Chemist, Cereal Chemistry Inc., St. Paul pp 1–78

    Google Scholar 

  • Thiellement H, Bahrman N, Damerval C, Plomion C, Rossignol M, Santoni V, de Vienne D, Zivy M (1999) Proteomics for genetic and physiological studies in plants. Electrophoresis 20:2013–2026

    Article  PubMed  CAS  Google Scholar 

  • Timothy JS, Isaiah TA (2000) Do more complex organisms have a greater proportion of membrane proteins in their genomes? Proteins 39:417–420

    Article  PubMed  Google Scholar 

  • Williams KL (1999) Genomes and proteomes: towards a multidimensional view of biology. Electrophoresis 20:678–688

    Article  PubMed  CAS  Google Scholar 

  • Williams KL, Hochstrasser DF (1997) Introduction to the proteome. In: Wilkins MR, Williams KL, Appel RD, Hochstrasser DF (eds) Proteome research: new frontiers in functional genomics. Springer, Berlin Heidelberg New York, pp 1–12

    Google Scholar 

  • Wong JH, Cai N, Balmer Y, Tanaka CK, Vensel WH, Hurkman WJ, Buchanan BB (2004) Thioredoxin targets of developing wheat seeds identified by complementary proteomic approaches. Phytochemistry 65:1629–1640

    Article  PubMed  CAS  Google Scholar 

  • Wu G, Wilen RW, Robertson AJ, Gusta LV (1999) Isolation, chromosomal localisation, and differential expression of mitochondrial manganese superoxide dismutase and chloroplastic copper/zinc superoxide dismutase genes in wheat. Plant Physiol 120(2):513–520

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

This research has been facilitated by access to the Australian Proteome Analysis Facility (APAF) established under the Australian Government’s Major National Research Facilities Program. This study was largely funded by the Value Added Wheat CRC, and Yunxian Mak is the recipient of a Value Added Wheat CRC Post-Graduate Scholarship.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Yunxian Mak.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mak, Y., Skylas, D.J., Willows, R. et al. A proteomic approach to the identification and characterisation of protein composition in wheat germ. Funct Integr Genomics 6, 322–337 (2006). https://doi.org/10.1007/s10142-005-0018-8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10142-005-0018-8

Keywords

Navigation