Partial purification of cytolytic venom proteins from the box jellyfish, Chironex fleckeri.
Venom proteins from the nematocysts of Chironex fleckeri were fractionated by size-exclusion and cation-exchange chromatography. Using sheep erythrocyte haemolysis as an indicator of cytolytic activity, two major cytolysins, with native molecular masses of approximately 370 and 145kDa, and one minor cytolysin ( approximately 70kDa) were isolated. SDS-PAGE and western blot protein profiles revealed that the 370kDa haemolysin is composed of CfTX-1 and CfTX-2 subunits ( approximately 43 and 45kDa, respectively); the most abundant proteins found in C. fleckeri nematocyst extracts. The 145kDa haemolysin predominately contains two other major proteins ( approximately 39 and 41kDa), which are not antigenic towards commercially available box jellyfish antivenom or rabbit polyclonal antibodies raised against whole C. fleckeri nematocyst extracts or CfTX-1 and -2. The kinetics of CfTX-1 and -2 haemolytic activities are temperature dependent and characterised by a pre-lytic lag phase ( approximately 6-7min) prior to initiation of haemolysis. Significant amino acid sequence homology between the CfTX proteins and other box jellyfish toxins suggest that CfTX-1 and -2 may also be lethal and dermonecrotic. Therefore, further in vivo and in vitro studies are required to investigate the potential roles of CfTX-1 and -2 in the lethal effects of C. fleckeri venom.
Brinkman
Diane
D
Department of Biochemistry and Molecular Biology, School of Pharmacy and Molecular Sciences, James Cook University, Townsville, Qld 4811, Australia. d.brinkman@aims.gov.au
Burnell
James
J
D000362:Agar / Q000737:chemistry; D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003064:Cnidarian Venoms / Q000737:chemistry* / Q000378:metabolism; D037462:Cubozoa / Q000378:metabolism*; D003603:Cytotoxins / Q000737:chemistry* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D006460:Hemolysin Proteins / Q000737:chemistry / Q000302:isolation & purification; D006461:Hemolysis; D011506:Proteins / Q000737:chemistry*; D040901:Proteomics
D003064:Cnidarian Venoms; D003603:Cytotoxins; D006460:Hemolysin Proteins; D011506:Proteins; D000362:Agar