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Identification of nuclear proteins of small cell lung cancer cell line H82: An improved procedure for the analysis of silver‐stained proteins
Author(s) -
González Luis Javier,
CastellanosSerra Lila,
Badock Volker,
Díaz Maylín,
Moro Alejandro,
Perea Silvio,
Santos Alicia,
PazLago Dalila,
Otto Albrecht,
Müller EvaChristina,
Kostka Susanne,
WittmannLiebold Brigitte,
Padrón Gabriel
Publication year - 2003
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/elps.200390020
Subject(s) - deamidation , mass spectrometry , proteome , asparagine , matrix assisted laser desorption/ionization , chemistry , electrospray ionization , chromatography , protein mass spectrometry , peptide mass fingerprinting , proteomics , microbiology and biotechnology , biochemistry , biology , amino acid , desorption , organic chemistry , adsorption , gene , enzyme
An efficient method for digestion and extraction of proteolytic peptides from silver‐stained proteins was applied to the characterization of nuclear proteins from the small cell lung cancer H82 (ATCC HTB 175) cell line previously separated by high‐resolution large format two‐dimensional gel electrophoresis. From 68 spots, evenly distributed on the gel area and representing a wide range of spot intensities, 63 (92%) were successfully identified by matrix‐assisted laser desorption/ionization (MALDI) or electrospray ionozation‐mass spectrometry (ESI‐MS). In five cases where the identification was not possible, the presence of an intense background apparently due to the leakage of polymers from the microtubes or other plastics, was detected. Extensive analysis of peptide sequences by ESI MS/MS experiments allowed the identification of post‐translational modifications, such as acetylation, phosphorylation, deamidation of asparagine residues and the presence of isoaspartic acid. A new protein variant not reported in sequence databases was also detected.