Akademik Veri Yönetim Sistemi
KAFKAS UNIVERSITESI VETERINER FAKULTESI DERGISI, cilt.14, sa.2, ss.179-184, 2008 (SCI-Expanded)
The aim of this study was to detect the glycosylated proteins of Helicobacter pylori (H. pylori) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and two dimensional gel electrophoresis (2-DE). Protein analysis of the H. pylori strains was performed using denaturing 8% SDS-PAGE. H. pylori 26695 cell proteins were also separated by 2-DE into hundreds of spots. Separated proteins of H. pylori strains by SDS-PAGE and 2-DE were transferred onto the Polyvinylidene Fluoride (PVDF) membrane by semi-dry blotting. Detection of glycosylated proteins of the protein bands or spots on blotted membranes were determined by overlay reactions with Digoxigenin (DIG)-Glycan Detection and Differentation kits (Roche Diagnostics, Germany). The Roche DIG-Glycan Differentiation Kit including peanut agglutinin (PNA) lectin was utilized to further characterize the glycosidic modifications. Analysis of protein bands on a blotted membrane with DIG Glycan Detection kit after SDS-PAGE analysis gave the general pattern of glycosylated proteins of H. pylori; interestingly PA4, PR20 and P12 strains of H. pylori gave the different patterns of protein glycosylation on 8% polyacrilamide gels. Moreover, a glycosylated protein band (similar to 54 kDa) was also detected dominantly on the outer membrane part of H. pylori. 2-DE analysis of H. pylori proteins showed about twelve clear spots indicating the O-glycosidically linked carbohydrate chains (galactose-beta(1-3)-N-acetylgalactosamine) determined by PNA lectin staining of the blotted membrane. Obtained results suggest the presence of some potentially glycosylated proteins in H. pylori.