Biologically active polypeptides from the tropical sea anemone Radianthus macrodactylus

• Published in: Toxicon (Oxford) Vol. 40; no. 8; pp. 1197 - 1217
• Main Authors: Monastyrnaya, Margarita M, Zykova, Tatyana A, Apalikova, Olga V, Shwets, Tatyana V, Kozlovskaya, Emma P
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.08.2002
• Subjects: Acetone; Actinoporins; Amino Acid Sequence; Amino Acids - analysis; Animals; Chromatography, Affinity; Chromatography, Gel; Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; Cnidarian Venoms - analysis; Cytolysins; Cytotoxins - chemistry; Cytotoxins - isolation & purification; Cytotoxins - toxicity; Electrophoresis, Polyacrylamide Gel; Hemolysis - drug effects; Hemolytic; Histamine Antagonists - pharmacology; Histaminolytic activity; Humans; In Vitro Techniques; Isoelectric Focusing; Lethal Dose 50; Lipids - blood; Marine; Molecular Sequence Data; Molecular Weight; Peptides - chemistry; Peptides - isolation & purification; Peptides - toxicity; Phospholipases A - metabolism; polypeptides; Proteins - analysis; Radianthus macrodactylus; Solvents; Sphingomyelin; Trypsin inhibitors; Trypsin Inhibitors - chemistry; Trypsin Inhibitors - isolation & purification; Trypsin Inhibitors - pharmacology; Trypsin inhibitory activity; Tumor Cells, Cultured; Trypsin inhibitors; Hemolytic; Histaminolytic activity; Trypsin inhibitory activity; Sphingomyelin; Actinoporins; Cytolysins
• ISSN: 0041-0101; 1879-3150
• DOI: 10.1016/S0041-0101(02)00139-3

Some biologically active polypeptides, three high and two low molecular weight cytolysins and four trypsin inhibitors were isolated from the sea anemone Radianthus macrodactylus and characterized. The purification steps involved acetone precipitation, gel filtration, ion-exchange, and affinity chromatography, and ion-exchange and reverse-phase HPLC. The relative molecular weight of high molecular weight Radianthus cytolysins named according to their N-terminal amino acids RTX-A (Ala), RTX-S (Ser) and RTX-G (Gly) was about 20,000. The isoelectric points were 9.8 for RTX-A and RTX-S, and 10.5 for RTX-G. The hemolytic activities of RTX-A, RTX-S and RTX-G were 3.5×10 4, 5.0×10 4, and 1.0×10 4 HU/mg, respectively, and were inhibited by sphingomyelin. The N-terminal amino acid sequence of RTX-A was determined as ALAGAIIAGAGLGLKILIEVLGEG–VKVKI–. Molecular weight of low molecular weight Radianthus cytolysins RmI, RmII, and of one trypsin inhibitor InI were 5100, 6100 and 7100, respectively. Isoelectric points for RmI and RmII were 9.2 and 9.3. Their hemolytic activity worked out 25 and 20 HU/mg, and was not inhibited by sphingomyelin. Toxicity of RmI and RmII was assessed by their histaminolytic activity. Amino acid composition of RmI and RmII was similar to that of tealiatoxin, histaminolytic cytolysin from the sea anemone Tealia felina.