Purification and partial characterization of the phospholipase A2 and co-lytic factor from sea anemone (Aiptasia pallida) nematocyst venom.
Functional nematocysts of one specific morphological class, the penetrant microbasic mastigophores, were isolated from the sea anemone, Aiptasia pallida. These nematocysts contain a multicomponent venom composed of several proteins, including those with neurotoxic, hemolytic, and lethal activities. Hemolytic activity is produced by at least three synergistic venom proteins. One of these proteins is identified as a phospholipase A2 (EC 3.1.1.4) which exists in two isozymic forms, alpha and beta, with molecular weights of 45,000 and 43,000, respectively. The beta isozyme has been purified to homogeneity. It is a single-chained glycoprotein with an isoelectric point (pI) of 8.8 and represents 70% of the phospholipase activity of the venom. The activity of the beta isozyme is relatively labile and is inactivated by 3.5 M urea or by heating at 45 degrees C. It is most stable at pH 4.0 and loses 50% of its activity at pH values below 3.5 and above 8.0. A second venom protein has also been purified. It is essential for the hemolytic activity of the venom and is termed co-lytic factor (CLF). It is a monomeric glycoprotein having a pI of 4.5. CLF has a molecular weight of approximately 98,000, a sedimentation coefficient of 4.8 S, and is prolate in shape, having a frictional ratio of about 1.6. CLF constitutes about 1.25% of the total venom protein and is assayed by reversing fatty acid inhibition of the venom hemolysis activity.
Grotendorst
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Department of Cell Biology and Anatomy, University of Miami School of Medicine, FL 33136, USA.
Hessinger
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DA
D000818:Animals; D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003064:Cnidarian Venoms / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000187:drug effects; D019791:Guanidine / Q000737:chemistry; D006460:Hemolysin Proteins / Q000032:analysis / Q000302:isolation & purification* / Q000494:pharmacology; D005779:Immunodiffusion; D007526:Isoelectric Point; D008387:Marine Toxins / Q000032:analysis / Q000302:isolation & purification* / Q000494:pharmacology; D008970:Molecular Weight; D010741:Phospholipases A / Q000032:analysis / Q000302:isolation & purification* / Q000494:pharmacology; D054467:Phospholipases A2; D051381:Rats; D012615:Sea Anemones; D013045:Species Specificity
D003064:Cnidarian Venoms; D006460:Hemolysin Proteins; D008387:Marine Toxins; D010741:Phospholipases A; D054467:Phospholipases A2; D019791:Guanidine