Strategic shotgun proteomics approach for efficient construction of an expression map of targeted protein families in hepatoma cell lines.

摘要: An expression map of the most abundant proteins in human hepatoma HepG2 cells was established by a combination complementary shotgun proteomics approaches. Two-dimensional liquid chromatography (LC)-nano electrospray ionization (ESI) tandem mass spectrometry (MS/MS) as well one-dimensional LC-matrix-assisted laser desorption/ionization MS/MS were evaluated and shown that additional separation introduced at peptide level not efficient simple prefractionation protein extracts extending range total number identified. Direct LC-nanoESI analyses peptides from solubilized fraction excised gel bands sodium dodecyl sulfate-polyacrylamide electrophoresis fractionated insolubilized afforded best construction nonredundant cell map. Compiling data multiple variations rapid is nonetheless useful to increase sequence coverage confidence hits especially for those identified primarily single or two matches. While returned hit score general reflects abundance respective proteins, it reliable index differential expression. Using another closely related Hep3B comparative basis, 16 with more than two-fold difference defined spot intensity two-dimensional analysis which notably include members heat shock (Hsp) heterogeneous nuclear ribonucleoprotein (hnRPN) families. The observed higher hnRNP A2/B1 Hsp90 led search reported functional roles mediated concert both these multifunctional cellular chaperones. In agreement proposed model telomerase telomere bound promoting their interactions, obtained demonstrated could be correlated longer telomeric length tumorigenic Hep3B. This biological significance constitutes basis further delineation dynamic interactions modifications families how proteomic investigation mutually substantiated productive cycle hypothesis pattern driven research.