Proteome analysis of Saccharomyces cerevisiae: a methodological outline.
Proteome analysis offers a unique means of identifying important proteins, characterizing their modifications and beginning to describe their function. This is achieved through the combination of two technologies: protein separation and selection by two-dimensional gel electrophoresis, and protein identification and characterization by mass spectrometry. This methodological outline sketches the strengths and weaknesses of the two central technologies used, and provides both practical tips and the theoretical background for their utilization. One application of these technologies is illustrated by the characterization of genes, revealed by sequencing, but which have no--or only weak homology--to any other known genes. Other applications, for example the identification of protein markers for particular human diseases, are only referred to. The aim of the article is thus to provide the basis for a sound understanding of the full potential and limitations of proteome analysis.
Fey
S J
SJ
Centre for Proteome Analysis in Life Sciences, International Science Park Odense, Denmark.
Nawrocki
A
A
Larsen
M R
MR
Görg
A
A
Roepstorff
P
P
Skews
G N
GN
Williams
R
R
Larsen
P M
PM
D016208:Databases, Factual; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005656:Fungal Proteins / Q000235:genetics* / Q000302:isolation & purification*; D015870:Gene Expression; D016681:Genome, Fungal; D006801:Humans; D006863:Hydrogen-Ion Concentration; D013058:Mass Spectrometry / Q000379:methods; D009154:Mutation; D010449:Peptide Mapping / Q000379:methods; D012441:Saccharomyces cerevisiae / Q000737:chemistry* / Q000235:genetics*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D005656:Fungal Proteins