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

• Published in: Functional & integrative genomics Vol. 6; no. 4; pp. 322 - 337
• Main Authors: Mak, Yunxian, Skylas, Daniel J, Willows, Robert, Connolly, Angela, Cordwell, Stuart J, Wrigley, Colin W, Sharp, Peter J, Copeland, Les
• Format: Journal Article
• Language: English
• Published: Heidelberg Berlin/Heidelberg : Springer-Verlag 01.10.2006; Springer
• Subjects: Analytical, structural and metabolic biochemistry; Biological and medical sciences; Electrophoresis, Polyacrylamide Gel - methods; Fundamental and applied biological sciences. Psychology; Germination; Mass Spectrometry - methods; Miscellaneous; Molecular and cellular biology; Molecular genetics; Peptide Mapping - methods; Plant Proteins - analysis; Proteins; Proteome - analysis; Proteomics - methods; Seeds - chemistry; Triticum - chemistry; Triticum aestivum; Monocotyledones; Embryo; Peptides; Nutrition; Enzyme; Germination; Proteome; Identification; Cereal crop; Protein; Gramineae; Fingerprint method; Angiospermae; Proteomics; Database; Spermatophyta; Two dimensional electrophoresis; Oxidoreductases; Metabolic enzymes; Aminoacid sequence; Triticum aestivum; Cultivar
• ISSN: 1438-793X; 1438-7948
• DOI: 10.1007/s10142-005-0018-8

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.