Venom Proteome of the Box Jellyfish Chironex fleckeri

Abstract

The nematocyst is a complex intracellular structure unique to Cnidaria. When triggered to discharge, the nematocyst explosively releases a long spiny, tubule that delivers an often highly venomous mixture of components. The box jellyfish, Chironex fleckeri, produces exceptionally potent and rapid-acting venom and its stings to humans cause severe localized and systemic effects that are potentially life-threatening. In an effort to identify toxins that could be responsible for the serious health effects caused by C. fleckeri and related species, we used a proteomic approach to profile the protein components of C. fleckeri venom. Collectively, 61 proteins were identified, including toxins and proteins important for nematocyte development and nematocyst formation (nematogenesis). The most abundant toxins identified were isoforms of a taxonomically restricted family of potent cnidarian proteins. These toxins are associated with cytolytic, nociceptive, inflammatory, dermonecrotic and lethal properties and expansion of this important protein family goes some way to explaining the destructive and potentially fatal effects of C. fleckeri venom. Venom proteins and their post-translational modifications (PTMs) were further characterized using toxin-specific antibodies and phosphoprotein/glycoprotein-specific stains. Results indicated that glycosylation is a common PTM of the toxin family while a lack of cross-reactivity by toxin-specific antibodies infers there is significant divergence in structure and possibly function among family members. This study provides insight into the depth and diversity of protein toxins produced by harmful box jellyfish and represents the first description of a cubozoan jellyfish venom proteome.

Chironex fleckeri (Box Jellyfish) Venom Proteins

“…The column was calibrated using molecular mass standards (GE Healthcare/Sigma), including blue dextran (Ͼ2000 kDa), apoferritin (440 kDa), ␤-amylase (200 kDa), alcohol dehydrogenase (150 kDa), bovine serum albumen (66 kDa), albumin (45 kDa), carbonic anhydrase (29 kDa), cytochrome c (12 kDa), and vitamin B 12 (1.4 kDa). Protein elution was monitored by UV detection (280 nm), and fractions (1 ml) were collected and retained on ice.…”

“…containing CfTX-A and -B were prepared and subjected to MS/MS as described previously (12,16). Briefly, tryptic fragments from in-gel digests were chromatographically separated on a Dionex Ultimate 3000 HPLC system using an Agilent Zorbax 300SB-C18 (3.5 m, 150 mm ϫ 75 m) column and a linear gradient of 0 -80% solvent B over 60 min and directly introduced into the source of a QSTAR Elite Hybrid MS/MS system (AB Sciex) operated in positive ion electrospray mode.…”

Bioactive toxins from stinging jellyfish

“…DNA and RNA sequencing have been conducted for Carukia barnesi, Malo kingi (Avila Soria, 2009), and Aurelia aurita. Putative toxins were identified by proteomic analysis of nematocysts of Chironex fleckeri (Brinkman et al, 2012), Olindias sambaquiensis (Weston et al, 2013), and Stomolophus meleagris (Li et al, 2014).…”

“…A proteomic study of the venom of Chironex fleckeri was published in 2012 (Brinkman et al, 2012). CfTX-1 and CfTX-2 were identified as well as homologues of the toxins CqTX-A from Chiropsalmus quadrigatus, CaTX-A from Carybdea alata, CrTX-A from Carybdea rastoni and Cytotoxin A isoform 1 from Malo kingi.…”

High-throughput expression of animal venom toxins in Escherichia coli to generate a large library of oxidized disulphide-reticulated peptides for drug discovery

“…However, although the use of venoms for drug discovery is rapidly emerging, it is still mostly an unrealized prospect due to recurrent technical bottlenecks that represent venom exploration. The advent of -omic techniques [5–7] lead to an explosion of the number of toxin sequences known, which are available for biomedical and biotechnological exploitation [8]. High-throughput production of these highly biologically-relevant molecules using recombinant technologies is currently the major limiting step between sequencing and biological screening and represents nowadays a considerable challenge.…”