title: Chironex fleckeri (box jellyfish) venom proteins: expansion of a cnidarian toxin family that elicits variable cytolytic and cardiovascular effects.
abstract: The box jellyfish Chironex fleckeri produces extremely potent and rapid-acting venom that is harmful to humans and lethal to prey. Here, we describe the characterization of two C. fleckeri venom proteins, CfTX-A (∼40 kDa) and CfTX-B (∼42 kDa), which were isolated from C. fleckeri venom using size exclusion chromatography and cation exchange chromatography. Full-length cDNA sequences encoding CfTX-A and -B and a third putative toxin, CfTX-Bt, were subsequently retrieved from a C. fleckeri tentacle cDNA library. Bioinformatic analyses revealed that the new toxins belong to a small family of potent cnidarian pore-forming toxins that includes two other C. fleckeri toxins, CfTX-1 and CfTX-2. Phylogenetic inferences from amino acid sequences of the toxin family grouped CfTX-A, -B, and -Bt in a separate clade from CfTX-1 and -2, suggesting that the C. fleckeri toxins have diversified structurally and functionally during evolution. Comparative bioactivity assays revealed that CfTX-1/2 (25 μg kg(-1)) caused profound effects on the cardiovascular system of anesthetized rats, whereas CfTX-A/B elicited only minor effects at the same dose. Conversely, the hemolytic activity of CfTX-A/B (HU50 = 5 ng ml(-1)) was at least 30 times greater than that of CfTX-1/2. Structural homology between the cubozoan toxins and insecticidal three-domain Cry toxins (δ-endotoxins) suggests that the toxins have a similar pore-forming mechanism of action involving α-helices of the N-terminal domain, whereas structural diversification among toxin members may modulate target specificity. Expansion of the cnidarian toxin family therefore provides new insights into the evolutionary diversification of box jellyfish toxins from a structural and functional perspective.
authors.affiliation: From the Australian Institute of Marine Science, P.M.B. No 3, Townsville Mail Centre, Townsville, Queensland 4810, Australia.
authors.forename: Diane L
authors.initials: DL
authors.lastname: Brinkman
authors.forename: Nicki
authors.initials: N
authors.lastname: Konstantakopoulos
authors.forename: Bernie V
authors.initials: BV
authors.lastname: McInerney
authors.forename: Jason
authors.initials: J
authors.lastname: Mulvenna
authors.forename: Jamie E
authors.initials: JE
authors.lastname: Seymour
authors.forename: Geoffrey K
authors.initials: GK
authors.lastname: Isbister
authors.forename: Wayne C
authors.initials: WC
authors.lastname: Hodgson
mesh_terms: D000595:Amino Acid Sequence; D000758:Anesthesia; D000818:Animals; D001483:Base Sequence; D001794:Blood Pressure / Q000187:drug effects; D015153:Blotting, Western; D002319:Cardiovascular System / Q000187:drug effects*; D016923:Cell Death / Q000187:drug effects; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003064:Cnidarian Venoms / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D018076:DNA, Complementary / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D006461:Hemolysis / Q000187:drug effects; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D010802:Phylogeny; D020033:Protein Isoforms / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D020539:Sequence Analysis, Protein; D012756:Sheep
chemical_list: D003064:Cnidarian Venoms; D018076:DNA, Complementary; D020033:Protein Isoforms; C524740:toxin-1, Chironex fleckeri; C524741:toxin-2, Chironex fleckeri
keywords: Bioinformatics; Cardiovascular; Cubozoa; Cytolytic; Endotoxin; Jellyfish Toxin; Nematocyst; Protein Purification; Toxins; Venom