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Studies of quantitative analysis of protein expression in Saccharomyces cerevisiae
Author(s) -
Yan Jun X.,
Sanchez JeanCharles,
Tonella Luisa,
Williams Keith L.,
Hochstrasser Denis F.
Publication year - 1999
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/(sici)1522-2683(19990101)20:4/5<738::aid-elps738>3.0.co;2-2
Subject(s) - proteome , saccharomyces cerevisiae , context (archaeology) , amino acid analysis , protein expression , amino acid , biochemistry , amino acid residue , biology , chemistry , staining , chromatography , microbiology and biotechnology , yeast , peptide sequence , genetics , gene , paleontology
In the present study amino acid analysis is applied to quantitation of Saccharomyces cerevisiae proteome expression. The quantitation levels obtained are compared to data using densitometric analysis of silver or amido black staining and to the theoretical expression level (codon bias) of the identified proteins determined from their amino acid analysis (AAA). The results show that relative volume ratio (%vol) using Melanie II is a better parameter for spot quantitation than relative optical density ratio (%OD), and amino black staining provides good linearity within the range 1—100 pmol protein. However, AAA shows that theoretical expression levels are not well correlated with actual protein expression level, although there is better correlation when isoforms of the expressed protein are identified and included. It is concluded that amino acid analysis provides accurate protein quantitation and has a continuing role in proteome studies in terms of the rapid and inexpensive quantitation of proteins displayed on proteome maps. We do however recognize that in the context of future clinical applications and large‐scale proteome discovery projects, quantitation and post‐translational modification need to be analyzed by “proteomatic” ( i.e. , proteome automatic bioinformatic analysis directly from the gel) techniques.