Partial purification and characterization of a hemolysin (CAH1) from Hawaiian box jellyfish (Carybdea alata) venom.
We have isolated and characterized a novel hemolytic protein from the venom of the Hawaiian box jellyfish (Carybdea alata). Hemolysis of sheep red blood cells was used to quantitate hemolytic potency of crude venom extracted from isolated nematocysts and venom after fractionation and purification procedures. Hemolytic activity of crude venom was reduced or lost after exposure to the proteolytic enzymes trypsin, collagenase and papain. The activity exhibited lectin-like properties in that hemolysis was inhibited by D-lactulose and certain other sugars. Activity was irreversibly lost after dialysis of crude venom against divalent-free, 20mM EDTA buffer; it was optimal in the presence of 10mM Ca2+ or Mg2+. Two chromatographic purification methods, size fractionation on Sephadex G-200 and anion exchange with quaternary ammonium, provided fractions in which hemolytic activity corresponded to the presence of a protein band with an apparent molecular weight of 42kDa by SDS-PAGE. We have designated this protein as CAH1. The N-terminal sequence of CAH1 was determined to be: XAADAXSTDIDD/GIIG.
Békésy Laboratory of Neurobiology, Pacific Biomedical Research Center, University of Hawaii at Manoa, Honolulu, HI 96822, USA.
J J
JJ
Chung
L A
LA
Ratnapala
I M
IM
Cooke
A A
AA
Yanagihara
D000818:Animals; D002241:Carbohydrates / Q000633:toxicity; D002412:Cations / Q000737:chemistry; D002852:Chromatography, Ion Exchange; D003064:Cnidarian Venoms / Q000737:chemistry* / Q000302:isolation & purification / Q000633:toxicity; D010450:Endopeptidases / Q000737:chemistry; D006460:Hemolysin Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000633:toxicity; D006461:Hemolysis / Q000187:drug effects; D066298:In Vitro Techniques; D007202:Indicators and Reagents; D011506:Proteins / Q000737:chemistry; D012756:Sheep; D013696:Temperature
D002241:Carbohydrates; D002412:Cations; D003064:Cnidarian Venoms; D006460:Hemolysin Proteins; D007202:Indicators and Reagents; D011506:Proteins; C430289:hemolysin CAH1; D010450:Endopeptidases