title: Database of two-dimensional polyacrylamide gel electrophoresis of proteins labeled with CyDye DIGE Fluor saturation dye.
abstract: CyDye DIGE Fluor saturation dye (saturation dye, GE Healthcare Amersham Biosciences) enables highly sensitive 2-D PAGE. As the dye reacts with all reduced cysteine thiols, 2-D PAGE can be performed with a lower amount of protein, compared with CyDye DIGE Fluor minimal dye (GE Healthcare Amersham Biosciences), the sensitivity of which is equivalent to that of silver staining. We constructed a 2-D map of the saturation dye-labeled proteins of a liver cancer cell line (HepG2) and identified by MS 92 proteins corresponding to 123 protein spots. Functional classification revealed that the identified proteins had chaperone, protein binding, nucleotide binding, metal ion binding, isomerase activity, and motor activity. The functional distribution and the cysteine contents of the proteins were similar to those in the most comprehensive 2-D database of hepatoma cells (Seow et al.., Electrophoresis 2000, 21, 1787-1813), where silver staining was used for protein visualization. Hierarchical clustering on the basis of the quantitative expression profiles of the 123 characterized spots labeled with two charge- and mass-matched saturation dyes (Cy3 and Cy5) discriminated between nine hepatocellular carcinoma cell lines and primary cultured hepatocytes from five individuals, suggesting the utility of saturation dye and our database for proteomic studies of liver cancer.
authors.affiliation: Cancer Proteomics Project, National Cancer Center Research Institute, Tokyo, Japan.
authors.forename: Kazuyasu
authors.initials: K
authors.lastname: Fujii
authors.forename: Tadashi
authors.initials: T
authors.lastname: Kondo
authors.forename: Hideki
authors.initials: H
authors.lastname: Yokoo
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Okano
authors.forename: Masayo
authors.initials: M
authors.lastname: Yamada
authors.forename: Tesshi
authors.initials: T
authors.lastname: Yamada
authors.forename: Keiji
authors.initials: K
authors.lastname: Iwatsuki
authors.forename: Setsuo
authors.initials: S
authors.lastname: Hirohashi
mesh_terms: D045744:Cell Line, Tumor; D002478:Cells, Cultured; D003545:Cysteine / Q000737:chemistry; D030562:Databases, Protein; D015180:Electrophoresis, Gel, Two-Dimensional; D005456:Fluorescent Dyes / Q000737:chemistry*; D020869:Gene Expression Profiling; D022781:Hepatocytes / Q000737:chemistry / Q000166:cytology; D006801:Humans; D008113:Liver Neoplasms; D008969:Molecular Sequence Data; D011506:Proteins / Q000737:chemistry* / Q000145:classification
chemical_list: D005456:Fluorescent Dyes; D011506:Proteins; D003545:Cysteine