Screening and identification of glycoproteins from Campylobacter jejuni NCTC 11168 based on specific affinity between lectins and glycoproteins

Abstract:

Abstract:［Objective］ To screen glycoproteins from Campylobacter jejuni NCTC 11168．［Methods］ We introduced a screening strategy based on specific affinity between lectins and glycoproteins． First，the specific affinity between Soybean Agglutinin (SBA) and glycoproteins in C． jejuni was confirmed by western blotting． Then，magnetic beads coating with Soybean Agglutinin (SBA) were used to capture the putative glycoproteins． Putative glycoproteins were competitively eluted by N-acetylgalactosamine and separated by two-dimensional gel electrophoresis． Spots in gels were further analyzed by mass spectrometry． ［Results］A total of 22 proteins were identified by mass spectrometric analysis，of which 5 have ever been identified to be glycoproteins． Others including Cj0633 have not been proved to be glycosylated so far．［Conclusion］The method could be used to screen glycoproteins in bacteria．