PMC

A, assassin bug P. plagipennis envenomating feeder cricket (A. domestica). B, extraction of venom from P. plagipennis using electrostimulation.

Proteins detected by LC-MS/MS of 2D SDS-PAGE spots and HPLC fractions showing abundant proteases, CUB-domain proteins, and heteropteran venom family 1 proteins. A, 2D SDS-polyacrylamide gel of crude venom, showing protein families identified by LC-MS/MS of gel spots. B, HPLC trace of venom fractionation, showing protein families identified by LC-MS/MS of collected fractions.

Evolution of CUB domain proteins. Sequence names in blue indicate P. plagipennis sequences determined in this study. A, alignment of CUB domain venom proteins from P. plagipennis and the anthocorid O. laevigatus () with the N-terminal portion (including CUB domain) of P. plagipennis venom S1 protease 8. B, phylogeny according to Bayesian inference showing support for monophyly of cimicomorphan venom CUB domain proteins (highlighted yellow). Anthocorid CUB domain protein, GBG01000009.1; crustacean CUB-S1 protease, AAK48894.1; beetle CUB-S1 protease, ENN74674.1; mosquito CUB-S1 protease, XP_001657965.1.

Conserved sequence features in redulysin proteins. A, amino acid sequence alignment of redulysins with the cytolytic domain of trialysin (AAL82381.1). The consensus sequence of the redulysin proteins is shown below. Lys residues are shown in bold black, positively charged residues in red, and hydrophobic residues in blue. Residues predicted by PSIPRED to form α-helices are highlighted in gray. B, helical wheel diagram of the predicted helical portion of the consensus of redulysin sequences showing clustering of Lys residues and hydrophobic residues on opposite sides of the helix.

Radiation of triabin/lipocalin superfamily in Reduviidae and allied insects calculated by Bayesian inference. Branches of Pristhesancus, Triatoma, and Rhodnius proteins are colored orange, blue, and pink, respectively; node labels indicate posterior probabilities. Accessions of sequences used in this analysis are as follows: putative BABP (Triatoma), D1MX91; putative nitrophorin (Triatoma), A0A023F6B8; Triafestin, AB292809.1; RPAI, JA76747.1; Triplatin, BAE96121.1; Infestilin, AAZ38958.1; Pallidipin, AAA30329.1; Dimiconin, BAI50848.1; Triabin, CAA56540.1; Procalin, AEM97970.1; Triatin, AAZ38956.1; ApoD-like (Halyomorpha), XP_014291570.1; Isoallergen 1 (Blatella), C3RWZ4; ApoD-like (Cimex), XP_014256007.1; Uncharacterized (Cimex), XP_014250080.1; Lazarillo-like (Cimex), XP_014249890; Lipocalin (Homo), CAA47889.1.

Evolution of Ptu1-like peptides isolated from P. plagipennis venom. Signal sequences predicted by SignalP4.1 are shown in lowercase; lines above the text show the disulfide connectivity for Ptu1. Sequence labels in blue indicate peptides detected in venom using proteomics. POI, phenoloxidase inhibitor; AMP, antimicrobial peptide. Node labels indicate posterior probabilities. Comparison sequences are remipede (S. tulumensis) agatoxin-like peptide (), assassin bug venom peptides Ado1 (P58608.1), Iob1 (P58609.1), and Ptu1 (P58606.1); bee (Apis mellifera) OCLP1 (H9KQJ7.1); ant (Trachymyrmex cornetzi) predicted protein (KYN16117.1); ant (Wasmannia auropunctata) conotoxin-like peptide (XP_011706926.1); fly (Bradysia hygida) salivary putative AMP1 (ABA26621.1); and Hemiptera (Bemisia tabaci) putative AMP knottin peptide Btk-1 (ABC40569.1).