The Dictyostelium discoideum proteome - The SWISS-2DPAGE database of the multicellular aggregate (slug)

Jun X. Yan, Luisa Tonella, Jean Charles Sanchez, Marc R. Wilkins, Nicolle H. Packer, Andrew A. Gooley, Denis F. Hochstrasser, Keith L. Williams*

*Corresponding author for this work

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

The cellular slime mold Dictyostelium discoideum is a eukaryotic microorganism which has developmental life stages attractive to the cell and molecular biologist. By displaying the two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) protein map of different developmental stages, the key molecules can be identified and characterised, allowing a detailed understanding of the D. discoideum proteome. Here we describe the preparation of reference gel of the D. discoideum multicellular aggregate, the slug. Proteins were separated by 2-D PAGE with immobilised pH gradients (pH 3.5-10) in the first dimension and sodium dodecyl sulfate (SDS)-PAGE in the second dimension. Micropreparative gels were electroblotted onto polyvinylidene difluoride (PVDF) membranes and 150 spots were visualised by amido black staining. Protein spots were excised and 31 were putatively identified by matching their amino acid composition, estimated isoelectric point (pI) and molecular weight (M(r)) against the SWISS-PROT database with the ExPASy AAcompID tool (http://expasy.hcuge. ch/ch2d/aacompi.html). A total of 25 proteins were identified by matching against database entries for D, discoideum, and another six by cross-species matching against database entries for Saccharomyces cerevisiae proteins. This map will be available in the SWISS-2DPAGE database.

Original languageEnglish
Pages (from-to)491-497
Number of pages7
JournalElectrophoresis
Volume18
Issue number3-4
DOIs
Publication statusPublished - Mar 1997

Keywords

  • amino acid analysis
  • dictyostelium discoideum
  • isoelectric point
  • molecular weight
  • proteome
  • two-dimensional polyacrylamide gel electrophoresis

Fingerprint Dive into the research topics of 'The <i>Dictyostelium discoideum</i> proteome - The SWISS-2DPAGE database of the multicellular aggregate (slug)'. Together they form a unique fingerprint.

Cite this