Abstract
The sea nettle (Chrysaora quinquecirrha), a venomous jellyfish, occurs in great abundance in the Chesapeake Bay and other coastal waters. This coelenterate inflicts injury to swimmers by injecting a toxin-coated thread into the skin. Recent investigations have demonstrated that soluble toxin released from nettle nematocysts is neurotoxic, myotoxic, and cardiotoxic, as well as capable of producing dermonecrosis and hemolysis (1-3). It has also been shown that Chrysaora toxin increased the sodium permeability of active membranes independent of ATPase or cyclic AMP (4). In order to understand the toxin's physiological action at the cellular level, its effect on rat liver mitochondria and lysosome preparations was studied.
Passage of the nematocyst fluid through Sephadex gels resulted in partial purification and separation of the lethal activity into at least 2 fractions (5). In the present investigation, isoelectric focusing was applied to further separate and define the active components.
Materials and Methods. Nematocyst suspensions (NS) were prepared from fresh Chrysaora tentacles according to previously described techniques (5). Sonic treatment of NS with 3 A for 90 sec ruptured the nematocysts and released the soluble toxins. This suspension was then centrifuged at 50,000g for 1 hr before the resultant supernatant (SU) was inoculated into a Sephadex G-200 gel column (30 cm height × 2 cm diam). Four milliliter fractions were collected and assayed for protein according to the method of Waddell (6). The ascending portion of the major peak in the protein curve of the eluate (FAS) was freeze-dried, resuspended in distilled water and inoculated into the isoelectric focusing apparatus (Model No. 8101, LKB Produkter AB, the Netherlands). Two to three watts were supplied to the focusing column for 2.5 days at 4°. At the conclusion of this run, serial 4 ml fractions of the sample within the column were collected.
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