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Proteome analysis of a human heptocellular carcinoma cell line, HCC‐M: An update
Author(s) -
Ou Keli,
Seow Teck Keong,
Liang Rosa C. M. Y.,
Ong ShaoEn,
Chung Maxey C. M.
Publication year - 2001
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200108)22:13<2804::aid-elps2804>3.0.co;2-g
Subject(s) - proteome , peptide mass fingerprinting , microbiology and biotechnology , chemistry , peptide , tandem mass spectrometry , spots , peptide sequence , mass spectrometry , human proteome project , proteomics , biology , biochemistry , chromatography , gene
Recently, we reported the proteome analysis of a human hepatocellular carcinoma cell line, HCC‐M ( Electrophoresis 2000, 21, 1787–1813), using two‐dimensional gel electrophoresis (2‐DE) and matrix assisted laser desorption/ionization‐time of flight‐mass spectrometry (MALDI‐TOF‐MS). From a total of 408 unique spots excised from the 2‐DE gel, 301 spots yielded good MALDI spectra. Out of these, 272 spots had matches returned from the database search leading to the identification of these proteins. Here, we report the results on the identification of the remaining 29 spots using nanoelectrospray ionization‐tandem mass spectrometry (nESI‐MS/MS). First, “peptide tag sequencing” was performed to obtain partial amino acid sequences of the peptides to search the SWISS‐PROT and NCBI nonredundant protein databases. Spots that were still not able to find any matches from the databases were subjected to de novo peptide sequencing. The tryptic peptide sequences were used to search for homologues in the protein and nucleotide databases with the NCBI Basic Local Alignment Search Tool (BLAST), which was essential for the characterization of novel or post‐translationally modified proteins. Using this approach, all the 29 spots were unambiguously identified. Among them, phosphotyrosyl phosphatase activator (PTPA), RNA‐binding protein regulatory subunit, replication protein A 32 kDa subunit (RP‐A) and N ‐acetylneuraminic acid phosphate synthase were reported to be cancer‐related proteins.*