Combined Peptidomic and Proteomic Analysis of Electrically Stimulated and Manually Dissected Venom from the South American Bullet Ant Paraponera clavata

Abstract

Ants have evolved venoms rich in peptides and proteins used for predation, defense, and communication. However, they remain extremely understudied due to the minimal amount of venom secreted by each ant. The present study investigated the differences in the proteome and peptidome of the venom from the bullet ant, Paraponera clavata. Venom samples were collected from a single colony either by manual venom gland dissection or by electrical stimulation and were compared using proteomic methods. Venom proteins were separated by 2D-PAGE and identified by nanoLC-ESI-QTOF MS/MS. Venom peptides were initially separated using C18 reversed-phase high-performance liquid chromatography, then analyzed by MALDI-TOF MS. The proteomic analysis revealed numerous proteins that could be assigned a biological function (total 94), mainly as toxins, or roles in cell regulation and transport. This investigation found that ca. 73% of the proteins were common to venoms collected by the two methods. The peptidomic analysis revealed a large number of peptides (total 309) but with <20% shared by the two collection methods. There was also a marked difference between venoms obtained by venom gland dissection from different ant colonies. These findings demonstrate the rich composition and variability of P. clavata venom.

Proteomic analysis of the venom of the predatory ant Pachycondyla striata (Hymenoptera: Formicidae)

“…This is the first characterization of the venom proteins of P. striata . Our data show that the venom of P. striata is highly diverse, similar to the venom of other ants that have been characterized (Aili et al., , ; Bouzid et al., ; Touchard et al., ). The combination of the three fractionation and analysis methods allowed the identification of 43 proteins in the venom of P. striata .…”

“…Ants employ venoms for a variety of purposes including predation and defense against predators and competitors, defense against microbial pathogens, social communication (Orivel & Dejean, ; Touchard et al., ), and as herbicides (Touchard et al., ). However, ant venoms remain almost unexplored in comparison with other animals (Aili et al., ; Rifflet et al., ; Touchard et al., ). The species of ants with the most well‐characterized venoms are the fire ant Solenopsis invicta and Solenopsis saevissima and Myrmecia pilosula (Santos et al., ; Touchard et al., ).…”

The Peptide Venom Composition of the Fierce Stinging Ant Tetraponera aethiops (Formicidae: Pseudomyrmecinae)

“…In the past, the investigation of ant venoms was greatly limited by the difficulty in gathering large amounts of venom due to the small size of these insects. However, the use of new techniques to collect venoms now permits large amounts of ant venom to be quickly obtained [12,13]. Furthermore, the use of multi-omics strategies to study ant venoms recently revealed the whole peptidome of three ant species belonging to the subfamilies Ponerinae, Myrmicinae, and Myrmeciinae, allowing a high-throughput identification of novel peptides.…”

Evaluation of Inter and Intraspecific Differences in the Venom Chemical Compositions of Polybia paulista Wasps and Ectatomma brunneum Ants Using FTIR-PAS

“…Studies analyzing only the protein portion of the P. paulista venom also show variation in the chemical composition between different colonies (Dias et al, 2014), as well as studies with ants that found variation in the venom composition of colonies of the same population (Aili et al, 2017), in which the authors suggested genetic polymorphism or small environmental variations between areas.…”