Proteomic characterization of wheat amyloplasts using identification of proteins by tandem mass spectrometry

Nancy L. Andon, Sarah Hollingworth, Antonius Koller, Andrew J. Greenland, John R. Yates, Paul A. Haynes*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

156 Citations (Scopus)

Abstract

We describe the initial characterization of the wheat amyloplast proteome, consisting of the identification and classification of 171 proteins. Whole amyloplasts and purified amyloplast membranes were prepared from wheat (Triticum aestivum). Protein extracts were examined by one-dimensional and two-dimensional electrophoresis, followed by high performance liquid chromatography-tandem mass spectrometry of separated proteins. Tandem mass spectrometry data of individual peptides was then searched by SEQUEST, using a database containing known protein sequences from both wheat and other homologous cereal crops. Using this approach we identified 108 proteins from whole amyloplasts and 63 proteins from purified amyloplast membranes. The majority of protein identifications were derived from protein sequences from cereal crops other than wheat, for which relatively little gene sequence data is available. The highest percentage of protein identifications obtained from any individual species was 46% of the total number of proteins identified, using sequence data found in our proprietary rice (Oryza sativa) genome database.

Original languageEnglish
Pages (from-to)1156-1168
Number of pages13
JournalProteomics
Volume2
Issue number9
DOIs
Publication statusPublished - 1 Sep 2002
Externally publishedYes

Keywords

  • Amyloplast
  • Plant
  • Proteome
  • Tandem mass spectrometry
  • Wheat

Fingerprint

Dive into the research topics of 'Proteomic characterization of wheat amyloplasts using identification of proteins by tandem mass spectrometry'. Together they form a unique fingerprint.

Cite this