PMID:
9150929
Authors:
Yan JX, Tonella L, Sanchez JC, Wilkins MR, Packer NH, Gooley AA,
Hochstrasser DF, Williams KL.
Title:
The Dictyostelium discoideum proteome--the SWISS-2DPAGE database of the
multicellular aggregate (slug).
Journal:
Electrophoresis. 1997 Mar-Apr;18(3-4):491-7. doi: 10.1002/elps.1150180325.
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.
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