Abstract
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 Polychrome-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 × 104 HU/mg, respectively. Exogenous sphingomyelin inhibits their action on the erythrocyte membrane. The N-terminal sequence of RTX-A was determined to be ALAGAIIA-GAGL/KGLKI/FLIEVLGEG-V/NKVKI-.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 648-655 |
| Number of pages | 8 |
| Journal | Russian Journal of Bioorganic Chemistry |
| Volume | 25 |
| Issue number | 10 |
| State | Published - Oct 1 1999 |
Keywords
- Cytolytic toxins
- Function
- Hemolytic activity
- Sea anemone
- Structure