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.
Absar
Nurul
N
Department of Biochemistry and Molecular Biology, University of Rajshahi, Rajshahi, 6205, Bangladesh. nabsar51@yahoo.com
Hasan
Sohel
S
Arisaka
F
F
D000595:Amino Acid Sequence; D000818:Animals; D029941:Fish Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000633:toxicity; D006023:Glycoproteins / Q000737:chemistry / Q000302:isolation & purification; D006385:Hemagglutination Inhibition Tests; D037241:Mannose-Binding Lectins / Q000737:chemistry* / Q000302:isolation & purification / Q000633:toxicity; D008934:Mitogens / Q000494:pharmacology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D051381:Rats; D016415:Sequence Alignment; D023721:Tetraodontiformes
D029941:Fish Proteins; D006023:Glycoproteins; D037241:Mannose-Binding Lectins; D008934:Mitogens