Purification, Characterization and N-terminal Sequences Alignment of a Mannose Specific Protein Purified from Potca Fish, Tetraodon patoca

A protein was isolated and purified from the ventral portion of the Potca fish, Tetraodon patoca. The method was accomplished by gel filtration of crude protein extract on Sephadex G-50 followed by Ion exchange chromatography on DEAE-cellulose and finally by affinity chromatography on ConA-Sepharose matrix. The molecular weight of the protein, determined by the gel filtration and SDS-PAGE was about 82,000 and 80,000 respectively, but 42,000 and 38,000 were indicated by SDS-PAGE in the presence of 2-mercaptoethanol. The protein agglutinated rat red blood cells and in a haptein-inhibition test, the protein was inhibited specifically by the d-mannose and mannose containing saccharides. The protein is glycoprotein with neutral sugar content of about 0.35%. The purified protein also showed strong cytotoxic effects, which was performed by brine shrimp lethality bioassay and histopathological examinations. The N-terminal amino acid sequences of both the subunits of the protein were also identified and used a blast search on N-terminal amino acid sequences of the subunits revealed that the protein showed significant homology with the homologous proteins in database.