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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

Item Type:Article
Title:Identification of nuclear proteins of small cell lung cancer cell line H82: An improved procedure for the analysis of silver-stained proteins
Creators Name:Gonzalez, L.J. and Castellanos-Serra, L. and Badock, V. and Diaz, M. and Moro, A. and Perea, S. and Santos, A. and Paz-Lago, D. and Otto, A. and Mueller, E.C. and Kostka, S. and Wittmann-Liebold, B. and Padron, G.
Abstract: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.
Keywords:In-Gel Digestion, Mass Spectrometry, Proteomics, Small-Cell Lung Cancer, Two-Dimensional Gel Electrophoresis
Source:Electrophoresis
ISSN:0173-0835
Publisher:Wiley (Germany)
Volume:24
Number:1-2
Page Range:237-252
Date:January 2003
Official Publication:https://doi.org/10.1002/elps.200390020
PubMed:View item in PubMed

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