[Isolation and characteristics of high molecular weight cytolysins from the sea anemone Radianthus macrodactylus].
Three cytolytic toxins (RTX: RTX-A, RTX-S, and RTX-G) were isolated from the sea anemone Radianthus macrodactylus and characterized. The purification scheme involved hydrophobic chromatography on Polychrom-1, batch-chromatography on CM-23 cellulose, gel filtration on Akrilex P-4, cation-exchange chromatography on CM-32 cellulose, and HPLC on an ion-exchange Ultropac TSK CM-3SW column and a reversed-phase Silasorb C18 column. The molecular masses of RTXs (ca. 20 kDa) were determined by SDS-PAGE in a density gradient of PAG. They are highly basic polypeptides (pI of 9.8 for RTX-A and RTX-S and 10.5 for RTX-G) containing similar amino acid compositions with a high content of basic and hydrophobic residues and the absence of Cys residues. The hemolytic activities of RTX-A, RTX-S, and RTX-G were determined to be 3.5, 5.0, and 1.0 x 10(4) HU/mg, respectively. Exogenous sphingomyelin inhibits their action on the erythrocyte membrane. The N-terminal sequence of RTX-A was determined to be ALAGAIIAGAGL/KGLKI/FLIEVLGEG--V/NKVKI-.
Monastyrnaia
M M
MM
Pacific Institute of Bioorganic Chemistry, Far-Eastern Division, Russian Academy of Sciences, Vladivostok, Russia. rita@piboc.marine.su
Zykova
T A
TA
Kozlovskaia
E P
EP
D000595:Amino Acid Sequence; D000818:Animals; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003603:Cytotoxins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D008969:Molecular Sequence Data; D012615:Sea Anemones / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
D003603:Cytotoxins