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Protein Glycosylation in the Gastric Pathogen Helicobacter pylori
| Content Provider | Semantic Scholar |
|---|---|
| Author | Naumowicz, Monika Kruszewski, Marcin Koty, Joanna Cechowska-Pasko, Marzanna |
| Copyright Year | 2017 |
| Abstract | Glycosylation was previously considered to be restricted to eukaryotes; however, through advances in analytical methods, there have been increasing reports of both O-linked and N-linked protein glycosylation pathways in bacteria, particularly amongst mucosal-associated pathogens. The purpose of this study was to detect the glycosylated proteins of Helicobacter pylori (H. pylori) separated by Sodium Dodecyl Sulfate Polyacrylamide Gel eEectrophoresis (SDS-PAGE) and Two Dimensional Gel Electrophoresis (2-DE). Protein analysis of the H. pylori strains was performed using denaturing 8% SDSPAGE. H. pylori 26695 cell proteins were also separated by 2-DE into hundreds of spots. Separated proteins of H. pylori strains by SDSPAGE 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). 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 (~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–β(1-3)-Nacetylgalactosamine) determined by PNA lectin staining of the blotted membrane. Obtained results of this study and current studies on the protein glycosylation in the gastric pathogen H. Pylori will be discussed. Effects of Fibronectins on Cell Morphology, Cell Attachment and Cell Spreading |
| File Format | PDF HTM / HTML |
| Alternate Webpage(s) | http://biochem.conference-site.com/archive/2017/proceedings/proceedings_ICBMB_2017.pdf |
| Language | English |
| Access Restriction | Open |
| Content Type | Text |
| Resource Type | Article |
