spider phylogeny; RTA clade; proteome analysis; venom composition; envenomation process; antimicrobial peptides; bioassays; mass spectrometry; enzymes; transcriptome analysis
Kuhn-Nentwig Lucia, Langenegger Nicolas, Heller Manfred, Koua Dominique, Nentwig Wolfgang (2019), The Dual Prey-Inactivation Strategy of Spiders—In-Depth Venomic Analysis of Cupiennius salei, in Toxins
, 11(3), 167-167.
NentwigWolfgang, PantiniPaolo, VetterRichard S. (2017), Distribution and medical aspects of Loxosceles rufescens, one of the most invasive spiders of the world (Araneae: Sicariidae), in Toxicon
, 132, 19-28.
Oldrati Vera, Koua Dominique, Allard Pierre-Marie, Hulo Nicolas, Arrell Miriam, Nentwig Wolfgang, Lisacek Frédérique, Wolfender Jean-Luc, Kuhn-Nentwig Lucia, Stöcklin Reto (2017), Peptidomic and transcriptomic profiling of four distinct spider venoms, in PLOS ONE
, 12(3), e0172966-e0172966.
Koua D, Kuhn-Nentwig L (2017), Spider neurotoxins, short linear cationic peptides and venom proteins classification improved by an automated competition between exhaustive profile HMM classifiers., in Toxins
, 9(245), 1-17.
Biner Olivier, Trachsel Christian, Moser Aline, Kopp Lukas, Langenegger Nicolas, Kämpfer Urs, von Ballmoos Christoph, Nentwig Wolfgang, Schürch Stefan, Schaller Johann, Kuhn-Nentwig Lucia (2015), Isolation, N-glycosylations and Function of a Hyaluronidase-Like Enzyme from the Venom of the Spider Cupiennius salei, in PLOS ONE
, 10(12), e0143963-e0143963.
Kuhn-Nentwig Lucia, Schaller Johann, Schürch Stefan, Nentwig Wolfgang (2015), Venom of Cupiennius salei (Ctenidae)
, Springer Netherlands, Dordrecht.
SummarySpider venoms consist of many components which mainly can be attributed to three major functional groups present in (probably) all spider venoms: low molecular mass compounds, cysteine-knotted peptides (the classical neurotoxins), and enzymes. Further functional groups (low molecular mass acylpolyamines, antimicrobial peptides and large neurotoxic proteins) are restricted to one or a few spider families. Within the evolution of spiders and their venom, numerous modifications, new developments and replacements of venom compounds occurred but their overall importance for the envenomation process and possible implications within the phylogeny of spiders are not well investigated. With this proposal, we will focus on the importance of (1) antimicrobial peptides and of (2) venom enzymes. (1) Cationic a-helical antimicrobial peptides destroy membranes and enhance thereby the effect of neurotoxins. So far, they are only known from species in a few lycosid-related families and from Zodariidae, both belonging to the RTA clade. Since antimicrobial peptides could speed up the envenomation process considerably, we hypothesize that they are a new invention (autapomorphy) within the RTA clade which represents the evolutionary youngest spider families. (2) There are several enzymes known from the venom of single spider species, either attacking the extracellular matrix or cell membranes and matrix proteins. These enzymes may act as “spreading factor” but experimental evidence for spider venoms is lacking. We assume that enzymes in spider venoms contribute considerably to the envenomation process but different spider families or family groups may have found different solutions.Our approach includes transcriptomic and proteomic studies of 50 spider species from 30 families. Antimicrobial peptide identification from different venoms will be done by specific cytolytic assays. Transcriptome (mRNA structure of antimicrobial peptides and neurotoxins) analysis will be done in combination with LC-MS/MS. Enzyme identification and characterization of three key enzymes will be done by specific enzyme/substrate assays. Sequence data of enzymes from C. salei will be obtained by a combination of N-terminal Edman sequencing of purified enzymes and mass spectrometry based identification using Peaks mass spectrometry software in combination with the venom gland cDNA library. With these obtained sequence data, different BLASTpn/p/x searches will be performed against different spider venom gland libraries, resulting in cDNA data of the three enzymes. These data will be confirmed by LC-MS/MS and mass spectrometry based identification of pretreated (reduced/alkylated and enzymatically digested) venom samples. We use Drosophila bioassays to quantify spreading effects of enzymes in dependency on coinjected neurotoxin CSTx-1 and cytolytically acting cupiennin 1a.Our goal is to explore the abundance and diversity of antimicrobial peptides within spider families, specifically to establish them as a new development of the RTA clade. The presence/absence data of enzymes in spider venoms and knowledge of their substrate specificity will give as insight into the importance of enzymes for the envenomation process. Both will allow us to draw some general conclusions on the level of functional aspects of major venom compound groups within spiders, but also, by sequence comparison, to investigate evolutionary dynamics between families or family groups.