Separation of Glycosylated OmpA-Like Proteins from Porphyromonas gingivalis and Tannerella forsythia.

Yukitaka Murakami, Keiji Nagano, Yoshiaki Hasegawa

Methods in molecular biology (Clifton, N.J.) 2021

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OmpA-like proteins located in the outer bacterial membrane are potential virulence factors from the major periodontal pathogens Porphyromonas gingivalis and Tannerella forsythia. Our previous studies have shown that OmpA-like proteins are glycosylated by O-linked N-acetylglucosamine (O-GlcNAc) and are strongly reactive to wheat germ agglutinin (WGA) lectin, which shows sugar specificity to GlcNAc. Utilizing this property, we have developed a separation method for OmpA-like proteins by affinity chromatography using WGA lectin-agarose. The purity of enriched native OmpA-like proteins were confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Coomassie Brilliant Blue (CBB) staining. More importantly, the purified OmpA-like proteins formed a unique trimeric structure keeping their bioactivity intact. In this chapter, we describe a detailed procedure to separate OmpA-like proteins, which may be used to further progress the biological studies of OmpA-like proteins.

Citation

Yukitaka Murakami, Keiji Nagano, Yoshiaki Hasegawa. Separation of Glycosylated OmpA-Like Proteins from Porphyromonas gingivalis and Tannerella forsythia. Methods in molecular biology (Clifton, N.J.). 2021;2210:143-155