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Two‐dimensional gel electrophoresis of gastric tissue in an alkaline p H range
Author(s) -
Kočevar Nina,
Grazio Snježana Frković,
Komel Radovan
Publication year - 2014
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201200574
Subject(s) - two dimensional gel electrophoresis , gel electrophoresis , chemistry , electrophoresis , range (aeronautics) , chromatography , microbiology and biotechnology , biochemistry , biology , proteomics , materials science , gene , composite material
2 DE in combination with MS has facilitated the discovery of several proteins with altered abundance in gastric cancer. While acidic and wide p H ranges have been widely investigated, analysis in the alkaline p H range has not been specifically performed in gastric cancer to date. In the present study, we initially optimized the 2 DE in alkaline pH range (pH 7–11) for gastric tissue samples. Using a modified lysis buffer, we analyzed pooled nontumor and tumor samples for proteins with altered abundance in gastric adenocarcinoma. We successfully identified 38 silver‐stained spots as 24 different proteins. Four of these were chosen for investigation with immunoblotting on individual paired samples to determine whether the changes seen in 2 DE represent the overall abundance of the protein or possibly only a single form. While mitochondrial trifunctional protein ( MTP ) subunits were decreased in 2 DE gels, immunoblotting identified their overall abundance as being differently dysregulated: in the gastric tumor samples, the MTP ‐α subunit was decreased, and the MTP ‐β subunit was increased. On the other hand, heterogenous nuclear ribonucleoprotein M and galectin‐4 were increased in the gastric tumor samples in both 2 DE and immunoblotting.