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In the venom of spiders, linear peptides (LPs), also called cytolytical or antimicrobial peptides, represent a largely neglected group of mostly membrane active substances that contribute in some spider species considerably to the killing power of spider venom. By next-generation sequencing venom gland transcriptome analysis, we investigated 48 spider species from 23 spider families and detected LPs in 20 species, belonging to five spider families (Ctenidae, Lycosidae, Oxyopidae, Pisauridae, and Zodariidae). The structural diversity is extraordinary high in some species: the lynx spider Oxyopes heterophthalmus contains 62 and the lycosid Pardosa palustris 60 different LPs. In total, we identified 524 linear peptide structures and some of them are in lycosids identical on amino acid level. LPs are mainly encoded in complex precursor structures in which, after the signal peptide and propeptide, 13 or more LPs (Hogna radiata) are connected by linkers. Besides Cupiennius species, also in Oxyopidae, posttranslational modifications of some precursor structures result in the formation of two-chain peptides. It is obvious that complex precursor structures represent a very suitable and fast method to produce a high number and a high diversity of bioactive LPs as economically as possible. At least in Lycosidae, Oxyopidae, and in the genus Cupiennius, LPs reach very high Transcripts Per Kilobase Million values, indicating functional importance within the envenomation process.
Lucia Kuhn-Nentwig; Heidi E. L. Lischer; Stano Pekár; Nicolas Langenegger; Maria J. Albo; Marco Isaia; Wolfgang Nentwig. Linear Peptides—A Combinatorial Innovation in the Venom of Some Modern Spiders. Frontiers in Molecular Biosciences 2021, 8, 1 .
AMA StyleLucia Kuhn-Nentwig, Heidi E. L. Lischer, Stano Pekár, Nicolas Langenegger, Maria J. Albo, Marco Isaia, Wolfgang Nentwig. Linear Peptides—A Combinatorial Innovation in the Venom of Some Modern Spiders. Frontiers in Molecular Biosciences. 2021; 8 ():1.
Chicago/Turabian StyleLucia Kuhn-Nentwig; Heidi E. L. Lischer; Stano Pekár; Nicolas Langenegger; Maria J. Albo; Marco Isaia; Wolfgang Nentwig. 2021. "Linear Peptides—A Combinatorial Innovation in the Venom of Some Modern Spiders." Frontiers in Molecular Biosciences 8, no. : 1.
Analysis of spider venom gland transcriptomes focuses on the identification of possible neurotoxins, proteins and enzymes. Here, the first comprehensive transcriptome analysis of cupiennins, small linear cationic peptides, also known as cytolytic or antimicrobial peptides, is reported from the venom gland transcriptome of Cupiennius salei by 454- and Illumina 3000 sequencing. Four transcript families with complex precursor structures are responsible for the expression of 179 linear peptides. Within the transcript families, after an anionic propeptide, cationic linear peptides are separated by anionic linkers, which are transcript family specific. The C-terminus of the transcript families is characterized by a linear peptide or truncated linkers with unknown function. A new identified posttranslational processing mechanism explains the presence of the two-chain CsTx-16 family in the venom. The high diversity of linear peptides in the venom of a spider and this unique synthesis process is at least genus specific as verified with Cupiennius getazi.
Lucia Kuhn-Nentwig. Complex precursor structures of cytolytic cupiennins identified in spider venom gland transcriptomes. Scientific Reports 2021, 11, 1 -15.
AMA StyleLucia Kuhn-Nentwig. Complex precursor structures of cytolytic cupiennins identified in spider venom gland transcriptomes. Scientific Reports. 2021; 11 (1):1-15.
Chicago/Turabian StyleLucia Kuhn-Nentwig. 2021. "Complex precursor structures of cytolytic cupiennins identified in spider venom gland transcriptomes." Scientific Reports 11, no. 1: 1-15.
The venom of Cupiennius salei is composed of dozens of neurotoxins, with most of them supposed to act on ion channels. Some insecticidal monomeric neurotoxins contain an α-helical part besides their inhibitor cystine knot (ICK) motif (type 1). Other neurotoxins have, besides the ICK motif, an α-helical part of an open loop, resulting in a heterodimeric structure (type 2). Due to their low toxicity, it is difficult to understand the existence of type 2 peptides. Here, we show with the voltage clamp technique in oocytes of Xenopus laevis that a combined application of structural type 1 and type 2 neurotoxins has a much more pronounced cytolytic effect than each of the toxins alone. In biotests with Drosophila melanogaster, the combined effect of both neurotoxins was enhanced by 2 to 3 log units when compared to the components alone. Electrophysiological measurements of a type 2 peptide at 18 ion channel types, expressed in Xenopus laevis oocytes, showed no effect. Microscale thermophoresis data indicate a monomeric/heterodimeric peptide complex formation, thus a direct interaction between type 1 and type 2 peptides, leading to cell death. In conclusion, peptide mergers between both neurotoxins are the main cause for the high cytolytic activity of Cupiennius salei venom.
Benjamin Clémençon; Lucia Kuhn-Nentwig; Nicolas Langenegger; Lukas Kopp; Steve Peigneur; Jan Tytgat; Wolfgang Nentwig; Benjamin P. Lüscher. Neurotoxin Merging: A Strategy Deployed by the Venom of the Spider Cupiennius salei to Potentiate Toxicity on Insects. Toxins 2020, 12, 250 .
AMA StyleBenjamin Clémençon, Lucia Kuhn-Nentwig, Nicolas Langenegger, Lukas Kopp, Steve Peigneur, Jan Tytgat, Wolfgang Nentwig, Benjamin P. Lüscher. Neurotoxin Merging: A Strategy Deployed by the Venom of the Spider Cupiennius salei to Potentiate Toxicity on Insects. Toxins. 2020; 12 (4):250.
Chicago/Turabian StyleBenjamin Clémençon; Lucia Kuhn-Nentwig; Nicolas Langenegger; Lukas Kopp; Steve Peigneur; Jan Tytgat; Wolfgang Nentwig; Benjamin P. Lüscher. 2020. "Neurotoxin Merging: A Strategy Deployed by the Venom of the Spider Cupiennius salei to Potentiate Toxicity on Insects." Toxins 12, no. 4: 250.
The venom of predators should be under strong selection pressure because it is a costly substance and prey may potentially become resistant. Particularly in prey-specialized predators, venom should be selected for its high efficiency against the focal prey. Very effective venom paralysis has been observed in specialized predators, such as spiders preying on dangerous prey. Here, we compared the toxicity of the venoms of two prey-specialized species, araneophagous Palpimanus sp. and myrmecophagous Zodarion nitidum, and their related generalist species. We injected different venom concentrations into two prey types—the prey preferred by a specialist and an alternative prey—and observed the mortality and the paralysis of the prey within 24 h. We found that the venoms of specialists were far more potent towards the preferred prey than alternative prey. The venoms of generalists were similarly potent towards both prey types. In addition, we tested the efficacy of two venom fractions (smaller and larger than 10 kDa) in araneophagous Palpimanus sp. Compounds larger than 10 kDa paralyzed both prey types, but smaller compounds (
Ondřej Michálek; Lucia Kuhn-Nentwig; Stano Pekár. High Specific Efficiency of Venom of Two Prey-Specialized Spiders. Toxins 2019, 11, 687 .
AMA StyleOndřej Michálek, Lucia Kuhn-Nentwig, Stano Pekár. High Specific Efficiency of Venom of Two Prey-Specialized Spiders. Toxins. 2019; 11 (12):687.
Chicago/Turabian StyleOndřej Michálek; Lucia Kuhn-Nentwig; Stano Pekár. 2019. "High Specific Efficiency of Venom of Two Prey-Specialized Spiders." Toxins 11, no. 12: 687.
This review gives an overview on the development of research on spider venoms with a focus on structure and function of venom components and techniques of analysis. Major venom component groups are small molecular mass compounds, antimicrobial (also called cytolytic, or cationic) peptides (only in some spider families), cysteine-rich (neurotoxic) peptides, and enzymes and proteins. Cysteine-rich peptides are reviewed with respect to various structural motifs, their targets (ion channels, membrane receptors), nomenclature, and molecular binding. We further describe the latest findings concerning the maturation of antimicrobial, and cysteine-rich peptides that are in most known cases expressed as propeptide-containing precursors. Today, venom research, increasingly employs transcriptomic and mass spectrometric techniques. Pros and cons of venom gland transcriptome analysis with Sanger, 454, and Illumina sequencing are discussed and an overview on so far published transcriptome studies is given. In this respect, we also discuss the only recently described cross contamination arising from multiplexing in Illumina sequencing and its possible impacts on venom studies. High throughput mass spectrometric analysis of venom proteomes (bottom-up, top-down) are reviewed.
Nicolas Langenegger; Wolfgang Nentwig; Lucia Kuhn-Nentwig. Spider Venom: Components, Modes of Action, and Novel Strategies in Transcriptomic and Proteomic Analyses. Toxins 2019, 11, 611 .
AMA StyleNicolas Langenegger, Wolfgang Nentwig, Lucia Kuhn-Nentwig. Spider Venom: Components, Modes of Action, and Novel Strategies in Transcriptomic and Proteomic Analyses. Toxins. 2019; 11 (10):611.
Chicago/Turabian StyleNicolas Langenegger; Wolfgang Nentwig; Lucia Kuhn-Nentwig. 2019. "Spider Venom: Components, Modes of Action, and Novel Strategies in Transcriptomic and Proteomic Analyses." Toxins 11, no. 10: 611.
Most knowledge of spider venom concerns neurotoxins acting on ion channels, whereas proteins and their significance for the envenomation process are neglected. The here presented comprehensive analysis of the venom gland transcriptome and proteome of Cupiennius salei focusses on proteins and cysteine-containing peptides and offers new insight into the structure and function of spider venom, here described as the dual prey-inactivation strategy. After venom injection, many enzymes and proteins, dominated by α-amylase, angiotensin-converting enzyme, and cysteine-rich secretory proteins, interact with main metabolic pathways, leading to a major disturbance of the cellular homeostasis. Hyaluronidase and cytolytic peptides destroy tissue and membranes, thus supporting the spread of other venom compounds. We detected 81 transcripts of neurotoxins from 13 peptide families, whereof two families comprise 93.7% of all cysteine-containing peptides. This raises the question of the importance of the other low-expressed peptide families. The identification of a venom gland-specific defensin-like peptide and an aga-toxin-like peptide in the hemocytes offers an important clue on the recruitment and neofunctionalization of body proteins and peptides as the origin of toxins.
Lucia Kuhn-Nentwig; Nicolas Langenegger; Manfred Heller; Dominique Koua; Wolfgang Nentwig. The Dual Prey-Inactivation Strategy of Spiders-In-Depth Venomic Analysis of Cupiennius salei. Toxins 2019, 11, 167 .
AMA StyleLucia Kuhn-Nentwig, Nicolas Langenegger, Manfred Heller, Dominique Koua, Wolfgang Nentwig. The Dual Prey-Inactivation Strategy of Spiders-In-Depth Venomic Analysis of Cupiennius salei. Toxins. 2019; 11 (3):167.
Chicago/Turabian StyleLucia Kuhn-Nentwig; Nicolas Langenegger; Manfred Heller; Dominique Koua; Wolfgang Nentwig. 2019. "The Dual Prey-Inactivation Strategy of Spiders-In-Depth Venomic Analysis of Cupiennius salei." Toxins 11, no. 3: 167.
Nicolas Langenegger; Dominique Koua; Stefan Schürch; Manfred Heller; Wolfgang Nentwig; Lucia Kuhn-Nentwig. A venom neurotoxin precursor processing protease from the spider Cupiennius salei. Toxicon 2019, 158, S40 .
AMA StyleNicolas Langenegger, Dominique Koua, Stefan Schürch, Manfred Heller, Wolfgang Nentwig, Lucia Kuhn-Nentwig. A venom neurotoxin precursor processing protease from the spider Cupiennius salei. Toxicon. 2019; 158 ():S40.
Chicago/Turabian StyleNicolas Langenegger; Dominique Koua; Stefan Schürch; Manfred Heller; Wolfgang Nentwig; Lucia Kuhn-Nentwig. 2019. "A venom neurotoxin precursor processing protease from the spider Cupiennius salei." Toxicon 158, no. : S40.
Spider venom neurotoxins and cytolytic peptides are expressed as elongated precursor peptides, which are post-translationally processed by proteases to yield the active mature peptides. The recognition motifs for these processing proteases, first published more than ten years ago, include the Processing Quadruplet Motif (PQM) and the inverted Processing Quadruplet Motif (iPQM). However, the identification of the relevant proteases was still pending. Here we describe the purification of a neurotoxin precursor processing protease from the venom of the spider Cupiennius salei. The chymotrypsinlike serine protease is a 28 kDa heterodimer with optimum activity at venom′s pH of 6.0. We designed multiple synthetic peptides mimicking the predicted cleavage sites of neurotoxin precursors. Using these peptides as substrates, we confirm the biochemical activity of the protease in propeptide removal from neurotoxin precursors by cleavage C-terminal of the PQM. Furthermore, the PQM protease also cleaves the iPQM relevant for heterodimerization of a subgroup of neurotoxins. An involvement in the maturing of cytolytic peptides is very likely, due to high similarity of present protease recognition motifs. Finally, bioinformatics analysis, identifying sequences of homolog proteins from 18 spiders of 9 families, demonstrate the wide distribution and importance of the isolated enzyme for spiders. In summary, we establish the first example of a PQM protease, essential for maturing of spider venom neurotoxins. In the future, the here described protease may be established as powerful tool for production strategies of recombinant toxic peptides, adapted to the maturing of spider venom toxins.
Nicolas Langenegger; Dominique Koua; Stefan Schürch; Manfred Heller; Wolfgang Nentwig; Lucia Kuhn-Nentwig. Identification of a precursor processing protease from the spider Cupiennius salei essential for venom neurotoxin maturation. Journal of Biological Chemistry 2018, 293, 2079 -2090.
AMA StyleNicolas Langenegger, Dominique Koua, Stefan Schürch, Manfred Heller, Wolfgang Nentwig, Lucia Kuhn-Nentwig. Identification of a precursor processing protease from the spider Cupiennius salei essential for venom neurotoxin maturation. Journal of Biological Chemistry. 2018; 293 (6):2079-2090.
Chicago/Turabian StyleNicolas Langenegger; Dominique Koua; Stefan Schürch; Manfred Heller; Wolfgang Nentwig; Lucia Kuhn-Nentwig. 2018. "Identification of a precursor processing protease from the spider Cupiennius salei essential for venom neurotoxin maturation." Journal of Biological Chemistry 293, no. 6: 2079-2090.
Spider venoms are rich cocktails of bioactive peptides, proteins, and enzymes that are being intensively investigated over the years. In order to provide a better comprehension of that richness, we propose a three-level family classification system for spider venom components. This classification is supported by an exhaustive set of 219 new profile hidden Markov models (HMMs) able to attribute a given peptide to its precise peptide type, family, and group. The proposed classification has the advantages of being totally independent from variable spider taxonomic names and can easily evolve. In addition to the new classifiers, we introduce and demonstrate the efficiency of hmmcompete, a new standalone tool that monitors HMM-based family classification and, after post-processing the result, reports the best classifier when multiple models produce significant scores towards given peptide queries. The combined used of hmmcompete and the new spider venom component-specific classifiers demonstrated 96% sensitivity to properly classify all known spider toxins from the UniProtKB database. These tools are timely regarding the important classification needs caused by the increasing number of peptides and proteins generated by transcriptomic projects.
Dominique Koua; Lucia Kuhn-Nentwig. Spider Neurotoxins, Short Linear Cationic Peptides and Venom Protein Classification Improved by an Automated Competition between Exhaustive Profile HMM Classifiers. Toxins 2017, 9, 245 .
AMA StyleDominique Koua, Lucia Kuhn-Nentwig. Spider Neurotoxins, Short Linear Cationic Peptides and Venom Protein Classification Improved by an Automated Competition between Exhaustive Profile HMM Classifiers. Toxins. 2017; 9 (8):245.
Chicago/Turabian StyleDominique Koua; Lucia Kuhn-Nentwig. 2017. "Spider Neurotoxins, Short Linear Cationic Peptides and Venom Protein Classification Improved by an Automated Competition between Exhaustive Profile HMM Classifiers." Toxins 9, no. 8: 245.
Hyaluronidases are important venom components acting as spreading factor of toxic compounds. In several studies this spreading effect was tested on vertebrate tissue. However, data about the spreading activity on invertebrates, the main prey organisms of spiders, are lacking. Here, a hyaluronidase-like enzyme was isolated from the venom of the spider Cupiennius salei. The amino acid sequence of the enzyme was determined by cDNA analysis of the venom gland transcriptome and confirmed by protein analysis. Two complex N-linked glycans akin to honey bee hyaluronidase glycosylations, were identified by tandem mass spectrometry. A C-terminal EGF-like domain was identified in spider hyaluronidase using InterPro. The spider hyaluronidase-like enzyme showed maximal activity at acidic pH, between 40–60°C, and 0.2 M KCl. Divalent ions did not enhance HA degradation activity, indicating that they are not recruited for catalysis. Besides hyaluronan, the enzyme degrades chondroitin sulfate A, whereas heparan sulfate and dermatan sulfate are not affected. The end products of hyaluronan degradation are tetramers, whereas chondroitin sulfate A is mainly degraded to hexamers. Identification of terminal N-acetylglucosamine or N-acetylgalactosamine at the reducing end of the oligomers identified the enzyme as an endo-β-N-acetyl-D-hexosaminidase hydrolase. The spreading effect of the hyaluronidase-like enzyme on invertebrate tissue was studied by coinjection of the enzyme with the Cupiennius salei main neurotoxin CsTx-1 into Drosophila flies. The enzyme significantly enhances the neurotoxic activity of CsTx-1. Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. This is interpreted as a loss of this enzyme and fits quite well the current phylogenetic idea on a more isolated position of these families and can perhaps be explained by specialized prey catching techniques.
Olivier Felix Biner; Christian Trachsel; Aline Moser; Lukas Kopp; Nicolas Langenegger; Urs Kämpfer; Christoph Von Ballmoos; Wolfgang Nentwig; Stefan Schürch; Johann Schaller; Lucia Kuhn-Nentwig. Isolation, N-glycosylations and Function of a Hyaluronidase-Like Enzyme from the Venom of the Spider Cupiennius salei. PLOS ONE 2015, 10, e0143963 .
AMA StyleOlivier Felix Biner, Christian Trachsel, Aline Moser, Lukas Kopp, Nicolas Langenegger, Urs Kämpfer, Christoph Von Ballmoos, Wolfgang Nentwig, Stefan Schürch, Johann Schaller, Lucia Kuhn-Nentwig. Isolation, N-glycosylations and Function of a Hyaluronidase-Like Enzyme from the Venom of the Spider Cupiennius salei. PLOS ONE. 2015; 10 (12):e0143963.
Chicago/Turabian StyleOlivier Felix Biner; Christian Trachsel; Aline Moser; Lukas Kopp; Nicolas Langenegger; Urs Kämpfer; Christoph Von Ballmoos; Wolfgang Nentwig; Stefan Schürch; Johann Schaller; Lucia Kuhn-Nentwig. 2015. "Isolation, N-glycosylations and Function of a Hyaluronidase-Like Enzyme from the Venom of the Spider Cupiennius salei." PLOS ONE 10, no. 12: e0143963.
More than half of all spider species hunt prey without a web. To successfully subdue their prey, they use adapted capture behaviour and efficient grasping mechanisms to interrupt the prey's locomotion, and to restrain it from escaping during the subsequent handling for final envenomation. In this study, we investigated how the prey capture behaviour of different lycosoid spider species is related to leg morphology and venom efficiency; using high speed videography, feeding experiments, stereomicroscopy, scanning electron microscopy and LD50 venom bioassays. We found that different species employed different techniques when grasping their prey and these differences strongly correlate with the distribution and size of hairy adhesive leg pads (so‐called scopulae on pro‐ and retrolateral parts of legs) and erectable spines, which act in a complementary way. Our results indicate that the grasping and handling behaviour and leg morphology is crucial in restricting the prey's movements. However, none of these traits is directly related with venom efficiency.
Benjamin Eggs; Jonas Wolff; Lucia Kuhn-Nentwig; Stanislav N. Gorb; Wolfgang Nentwig. Hunting Without a Web: How Lycosoid Spiders Subdue their Prey. Ethology 2015, 121, 1166 -1177.
AMA StyleBenjamin Eggs, Jonas Wolff, Lucia Kuhn-Nentwig, Stanislav N. Gorb, Wolfgang Nentwig. Hunting Without a Web: How Lycosoid Spiders Subdue their Prey. Ethology. 2015; 121 (12):1166-1177.
Chicago/Turabian StyleBenjamin Eggs; Jonas Wolff; Lucia Kuhn-Nentwig; Stanislav N. Gorb; Wolfgang Nentwig. 2015. "Hunting Without a Web: How Lycosoid Spiders Subdue their Prey." Ethology 121, no. 12: 1166-1177.
The venom of the ctenid spider Cupiennius salei contains a variety of low molecular mass compounds, many small cationic peptides, disulfiderich peptides, and several enzymes. Among the low molecular mass compounds are taurine, histamine, and citric acid. Some small cationic peptides act membranolytically and destroy prokaryotic as well as eukaryotic cells. More than 40 small cationic peptides (SCPs) have been identified and characterized so far, of which several exert strong cytolytic effects and enable other venom compounds to reach their targets. Most of the disulfiderich peptides follow the ICK motifand act as neurotoxins or support the activity of other neurotoxins as enhancers. More than a dozen different Cupiennius salei toxins (CsTx) can be distinguished so far. CsTx-1 is the most abundant and most insecticidal neurotoxin in the venom of C. salei. It is composed of two domains, an N-terminal part exhibiting the ICK motif, which inhibits L-type Ca2+ ion channels, and a highly cationic C-terminal tailadopting an a-helical conformation, which destroys cell membranes. Several CsTx peptides consisting of two chains, connected by two disulfide bonds, enhance in nontoxic concentrations the insecticidal activity of other toxins. Among the enzymes, a highly active hyaluronidase is crucial for destroying tissue and assisting other venom compounds to spread into the target organism. The combined effects of synergistic and enhancing interactions between these components enable C. salei to inject a minimum quantity of venom to achieve maximum toxicity, thus optimizing its venom investment
Lucia Gerda Kuhn-Nentwig; Johann Schaller; Stefan Schürch; Wolfgang Nentwig. Venom of Cupiennius salei (Ctenidae). Spider Venoms 2015, 1 -19.
AMA StyleLucia Gerda Kuhn-Nentwig, Johann Schaller, Stefan Schürch, Wolfgang Nentwig. Venom of Cupiennius salei (Ctenidae). Spider Venoms. 2015; ():1-19.
Chicago/Turabian StyleLucia Gerda Kuhn-Nentwig; Johann Schaller; Stefan Schürch; Wolfgang Nentwig. 2015. "Venom of Cupiennius salei (Ctenidae)." Spider Venoms , no. : 1-19.
The most abundant cell types in the hemolymph of Cupiennius salei are plasmatocytes (70–80%) and granulocytes (20–30%). Both cells differ in shape, cytochemical and transmission electron microscopy staining of their cytoplasma and granules. According to MALDI-IMS (matrix-assisted laser desorption ionization mass spectrometry imaging), granulocytes exhibit ctenidin 1 (9510 Da) and ctenidin 3 (9568 Da), SIBD-1 (8675 Da), and unknown peptides with masses of 2207 and 6239 Da. Plasmatocytes exhibit mainly a mass of 6908 Da. Unknown peptides with masses of 1546 and 1960 Da were detected in plasmatocytes and granulocytes. Transmission electron microscopy confirms the presence of two compounds in one granule and cytochemical staining (light microscopy) tends to support this view. Two further hemocyte types (cyanocytes containing hemocyanin and prehemocytes as stem cells) are only rarely detected in the hemolymph. These four hemocyte types constitute the cellular part of the spider immune system and this is discussed in view of arachnid hemocyte evolution
Lucia Kuhn-Nentwig; Lukas S. Kopp; Wolfgang Nentwig; Beat Haenni; Kathrin Streitberger; Stefan Schürch; Johann Schaller. Functional differentiation of spider hemocytes by light and transmission electron microscopy, and MALDI-MS-imaging. Developmental & Comparative Immunology 2014, 43, 59 -67.
AMA StyleLucia Kuhn-Nentwig, Lukas S. Kopp, Wolfgang Nentwig, Beat Haenni, Kathrin Streitberger, Stefan Schürch, Johann Schaller. Functional differentiation of spider hemocytes by light and transmission electron microscopy, and MALDI-MS-imaging. Developmental & Comparative Immunology. 2014; 43 (1):59-67.
Chicago/Turabian StyleLucia Kuhn-Nentwig; Lukas S. Kopp; Wolfgang Nentwig; Beat Haenni; Kathrin Streitberger; Stefan Schürch; Johann Schaller. 2014. "Functional differentiation of spider hemocytes by light and transmission electron microscopy, and MALDI-MS-imaging." Developmental & Comparative Immunology 43, no. 1: 59-67.
Cupiennins are small cationic α-helical peptides from the venom of the ctenid spider Cupiennius salei which are characterized by high bactericidal as well as hemolytic activities. To gain insight into the determinants responsible for the broad cytolytic activities, two analogues of cupiennin 1a with different N-terminal hydrophobicities were designed. The insecticidal, bactericidal and hemolytic activities of these analogues were assayed and compared to the native peptide. Specifically, substitution of two N-terminal Phe residues by Ala results in less pronounced insecticidal and cytolytic activity, whereas a substitution by Lys reduces strongly its bactericidal activity and completely diminishes its hemolytic activity up to very high tested concentrations. Biophysical analyses of peptide/bilayer membrane interactions point to distinct interactions of the analogues with lipid bilayers, and dependence upon membrane surface charge. Indeed, we find that lower hemolytic activity was correlated with less surface association of the analogues. In contrast, our data indicate that the reduced bactericidal activity of the two cupiennin 1a analogues likely correspond to greater bilayer-surface localization of the peptides. Overall, ultimate insertion and destruction of the host cell membrane is highly dependent on the presence of Phe-2 and Phe-6 (Cu 1a) or Leu-6 (Cu 2a) in the N-terminal sequences of native cupiennins.
Lucia Kuhn-Nentwig; Tania Sheynis; Sofiya Kolusheva; Wolfgang Nentwig; Raz Jelinek. N-terminal aromatic residues closely impact the cytolytic activity of cupiennin 1a, a major spider venom peptide. Toxicon 2013, 75, 177 -186.
AMA StyleLucia Kuhn-Nentwig, Tania Sheynis, Sofiya Kolusheva, Wolfgang Nentwig, Raz Jelinek. N-terminal aromatic residues closely impact the cytolytic activity of cupiennin 1a, a major spider venom peptide. Toxicon. 2013; 75 ():177-186.
Chicago/Turabian StyleLucia Kuhn-Nentwig; Tania Sheynis; Sofiya Kolusheva; Wolfgang Nentwig; Raz Jelinek. 2013. "N-terminal aromatic residues closely impact the cytolytic activity of cupiennin 1a, a major spider venom peptide." Toxicon 75, no. : 177-186.
The venom of Cupiennius salei is characterized by (1) a high diversity of cytolytic compounds (linear cytolytic peptides and a cysteine-rich peptide exhibiting two domains: the ICK motif and an α-helical cytolytically acting domain), (2) the neurotoxic activity of ion-channel inhibitors, (3) a highly active hyaluronidase and (4) synergistic interactions between many of these components. The combined effects of synergistic and enhancing interactions between various components enable Cupiennius salei to inject a maximum of toxicity with a minimum of venom quantity, thus optimizing its venom investment.
Lucia Gerda Kuhn-Nentwig; Wolfgang Nentwig. The Cytotoxic Mode of Action of the Venom of Cupiennius salei (Ctenidae). Spider Ecophysiology 2012, 217 -228.
AMA StyleLucia Gerda Kuhn-Nentwig, Wolfgang Nentwig. The Cytotoxic Mode of Action of the Venom of Cupiennius salei (Ctenidae). Spider Ecophysiology. 2012; ():217-228.
Chicago/Turabian StyleLucia Gerda Kuhn-Nentwig; Wolfgang Nentwig. 2012. "The Cytotoxic Mode of Action of the Venom of Cupiennius salei (Ctenidae)." Spider Ecophysiology , no. : 217-228.
CsTx-1, the main neurotoxic acting peptide in the venom of the spider Cupiennius salei, is composed of 74 amino acid residues, exhibits an inhibitory cysteine knot motif, and is further characterized by its highly cationic charged C terminus. Venom gland cDNA library analysis predicted a prepropeptide structure for CsTx-1 precursor. In the presence of trifluoroethanol, CsTx-1 and the long C-terminal part alone (CT1-long; Gly-45–Lys-74) exhibit an α-helical structure, as determined by CD measurements. CsTx-1 and CT1-long are insecticidal toward Drosophila flies and destroys Escherichia coli SBS 363 cells. CsTx-1 causes a stable and irreversible depolarization of insect larvae muscle cells and frog neuromuscular preparations, which seem to be receptor-independent. Furthermore, this membranolytic activity could be measured for Xenopus oocytes, in which CsTx-1 and CT1-long increase ion permeability non-specifically. These results support our assumption that the membranolytic activities of CsTx-1 are caused by its C-terminal tail, CT1-long. Together, CsTx-1 exhibits two different functions; as a neurotoxin it inhibits L-type Ca2+ channels, and as a membranolytic peptide it destroys a variety of prokaryotic and eukaryotic cell membranes. Such a dualism is discussed as an important new mechanism for the evolution of spider venomous peptides.
Lucia Kuhn-Nentwig; Irina M. Fedorova; Benjamin P. Lüscher; Lukas S. Kopp; Christian Trachsel; Johann Schaller; Xuan Lan Vu; Thomas Seebeck; Kathrin Streitberger; Wolfgang Nentwig; Erwin Sigel; Lev Magazanik. A Venom-derived Neurotoxin, CsTx-1, from the Spider Cupiennius salei Exhibits Cytolytic Activities. Journal of Biological Chemistry 2012, 287, 25640 -25649.
AMA StyleLucia Kuhn-Nentwig, Irina M. Fedorova, Benjamin P. Lüscher, Lukas S. Kopp, Christian Trachsel, Johann Schaller, Xuan Lan Vu, Thomas Seebeck, Kathrin Streitberger, Wolfgang Nentwig, Erwin Sigel, Lev Magazanik. A Venom-derived Neurotoxin, CsTx-1, from the Spider Cupiennius salei Exhibits Cytolytic Activities. Journal of Biological Chemistry. 2012; 287 (30):25640-25649.
Chicago/Turabian StyleLucia Kuhn-Nentwig; Irina M. Fedorova; Benjamin P. Lüscher; Lukas S. Kopp; Christian Trachsel; Johann Schaller; Xuan Lan Vu; Thomas Seebeck; Kathrin Streitberger; Wolfgang Nentwig; Erwin Sigel; Lev Magazanik. 2012. "A Venom-derived Neurotoxin, CsTx-1, from the Spider Cupiennius salei Exhibits Cytolytic Activities." Journal of Biological Chemistry 287, no. 30: 25640-25649.
The multicomponent venom of the spider Cupiennius salei was separated by three different chromatographic strategies to facilitate subsequent analysis of peptidic venom components by tandem mass spectrometry (MALDI‐TOF‐MS and ESI‐MS), Edman degradation and amino acid analysis: (a) desalting of the crude venom by RP‐HPLC only, (b) chromatographic separation of the crude venom into 42 fractions by RP‐HPLC, and (c) multidimensional purification of the crude venom by size exclusion and cation exchange chromatography and RP‐HPLC. A total of 286 components were identified in the venom of C. salei by mass spectrometry and the sequence of 49 new peptides was determined de novo by Edman degradation and tandem mass spectrometry; 30 were C‐terminally amidated. The novel peptides were assigned to two main groups: (a) short cationic peptides and (b) Cys‐containing peptides with the inhibitor cystine knot motif. Bioinformatics revealed a limited number of substantial similarities, namely with the peptides CpTx1 from the spider Cheiracantium punctorium and U3‐ctenitoxin‐Asp1a from the South American fishing spider (Ancylometes sp.) and with sequences from a Lycosa singoriensis venom gland transcriptome analysis. The results clearly indicate that the quality of the data is strongly dependent on the chosen separation strategy. The combination of orthogonal analytical methods efficiently excludes alkali ion and matrix adducts, provides indispensable information for an unambiguous identification of isomasses, and results in the most comprehensive repertoire of peptides identified in the venom of C. salei so far.
Christian Trachsel; Doreen Siegemund; Urs Kämpfer; Lukas S. Kopp; Claudia Bühr; Jonas Grossmann; Christoph Lüthi; Mónica Cunningham; Wolfgang Nentwig; Lucia Kuhn-Nentwig; Stefan Schürch; Johann Schaller. Multicomponent venom of the spider Cupiennius salei: a bioanalytical investigation applying different strategies. The FEBS Journal 2012, 279, 2683 -2694.
AMA StyleChristian Trachsel, Doreen Siegemund, Urs Kämpfer, Lukas S. Kopp, Claudia Bühr, Jonas Grossmann, Christoph Lüthi, Mónica Cunningham, Wolfgang Nentwig, Lucia Kuhn-Nentwig, Stefan Schürch, Johann Schaller. Multicomponent venom of the spider Cupiennius salei: a bioanalytical investigation applying different strategies. The FEBS Journal. 2012; 279 (15):2683-2694.
Chicago/Turabian StyleChristian Trachsel; Doreen Siegemund; Urs Kämpfer; Lukas S. Kopp; Claudia Bühr; Jonas Grossmann; Christoph Lüthi; Mónica Cunningham; Wolfgang Nentwig; Lucia Kuhn-Nentwig; Stefan Schürch; Johann Schaller. 2012. "Multicomponent venom of the spider Cupiennius salei: a bioanalytical investigation applying different strategies." The FEBS Journal 279, no. 15: 2683-2694.
Cupiennius salei single insulin‐like growth factor binding domain protein (SIBD‐1) is an 8.6 kDa Cys‐, Pro‐, and Gly‐rich protein, discovered in the hemocytes of the Central American hunting spider Cupiennius salei. SIBD‐1 exhibits high sequence similarity to the N‐terminal domain of the insulin‐like growth factor‐binding protein superfamily and has been reported to play an important role in the spider's immune system. Here, the recombinant expression and the elucidation of the three‐dimensional structure of recombinant SIBD‐1 and the characterization of the sugar moiety at Thr2 of native SIBD‐1 is described in detail. Proteins 2012;
Christian Trachsel; Christine Widmer; Urs Kämpfer; Claudia Bühr; Tommy Baumann; Lucia Kuhn-Nentwig; Stefan Schürch; Johann Schaller; Ulrich Baumann. Structural and biochemical characterization of native and recombinant single insulin-like growth factor-binding domain protein (SIBD-1) from the Central American Hunting Spider Cupiennius salei (Ctenidae). Proteins: Structure, Function, and Bioinformatics 2012, 80, 2323 -2329.
AMA StyleChristian Trachsel, Christine Widmer, Urs Kämpfer, Claudia Bühr, Tommy Baumann, Lucia Kuhn-Nentwig, Stefan Schürch, Johann Schaller, Ulrich Baumann. Structural and biochemical characterization of native and recombinant single insulin-like growth factor-binding domain protein (SIBD-1) from the Central American Hunting Spider Cupiennius salei (Ctenidae). Proteins: Structure, Function, and Bioinformatics. 2012; 80 (9):2323-2329.
Chicago/Turabian StyleChristian Trachsel; Christine Widmer; Urs Kämpfer; Claudia Bühr; Tommy Baumann; Lucia Kuhn-Nentwig; Stefan Schürch; Johann Schaller; Ulrich Baumann. 2012. "Structural and biochemical characterization of native and recombinant single insulin-like growth factor-binding domain protein (SIBD-1) from the Central American Hunting Spider Cupiennius salei (Ctenidae)." Proteins: Structure, Function, and Bioinformatics 80, no. 9: 2323-2329.
Cupiennius salei single insulin-like growth factor-binding domain protein (SIBD-1), which exhibits an IGFBP N-terminal domain-like profile, was identified in the hemocytes of the spider C. salei. SIBD-1 was purified by RP-HPLC and the sequence determined by a combination of Edman degradation and 5′–3′- RACE PCR. The peptide (8676.08 Da) is composed of 78 amino acids, contains six intrachain disulphide bridges and carries a modified Thr residue at position 2. SIBD-1 mRNA expression was detected by quantitative real-time PCR mainly in hemocytes, but also in the subesophageal nerve mass and muscle. After infection, the SIBD-1 content in the hemocytes decreases and, simultaneously, the temporal SIBD-1 expression seems to be down-regulated. Two further peptides, SIBD-2 and IGFBP-rP1, also exhibiting IGFBP N-terminal domain variants with unknown functions, were identified on cDNA level in spider hemocytes and venom glands. We conclude that SIBD-1 may play an important role in the immune system of spiders.
Lucia Kuhn-Nentwig; Carlo R. Largiadèr; Kathrin Streitberger; SathyaN Chandru; Tommy Baumann; Urs Kämpfer; Johann Schaller; Stefan Schürch; Wolfgang Nentwig. Purification, cDNA structure and biological significance of a single insulin-like growth factor-binding domain protein (SIBD-1) identified in the hemocytes of the spider Cupiennius salei. Insect Biochemistry and Molecular Biology 2011, 41, 891 -901.
AMA StyleLucia Kuhn-Nentwig, Carlo R. Largiadèr, Kathrin Streitberger, SathyaN Chandru, Tommy Baumann, Urs Kämpfer, Johann Schaller, Stefan Schürch, Wolfgang Nentwig. Purification, cDNA structure and biological significance of a single insulin-like growth factor-binding domain protein (SIBD-1) identified in the hemocytes of the spider Cupiennius salei. Insect Biochemistry and Molecular Biology. 2011; 41 (11):891-901.
Chicago/Turabian StyleLucia Kuhn-Nentwig; Carlo R. Largiadèr; Kathrin Streitberger; SathyaN Chandru; Tommy Baumann; Urs Kämpfer; Johann Schaller; Stefan Schürch; Wolfgang Nentwig. 2011. "Purification, cDNA structure and biological significance of a single insulin-like growth factor-binding domain protein (SIBD-1) identified in the hemocytes of the spider Cupiennius salei." Insect Biochemistry and Molecular Biology 41, no. 11: 891-901.
Three novel glycine-rich peptides, named ctenidin 1-3, with activity against the Gram-negative bacterium E. coli, were isolated and characterized from hemocytes of the spider Cupiennius salei. Ctenidins have a high glycine content (>70%), similarly to other glycine-rich peptides, the acanthoscurrins, from another spider, Acanthoscurria gomesiana. A combination of mass spectrometry, Edman degradation, and cDNA cloning revealed the presence of three isoforms of ctenidin, at least two of them originating from simple, intronless genes. The full-length sequences of the ctenidins consist of a 19 amino acid residues signal peptide followed by the mature peptides of 109, 119, or 120 amino acid residues. The mature peptides are post-translationally modified by the cleavage of one or two C-terminal cationic amino acid residue(s) and amidation of the newly created mature C-terminus. Tissue expression analysis revealed that ctenidins are constitutively expressed in hemocytes and to a small extent also in the subesophageal nerve mass
Tommy Baumann; Urs Kämpfer; Stefan Schürch; Johann Schaller; Carlo Largiader; Wolfgang Nentwig; Lucia Kuhn-Nentwig. Ctenidins: antimicrobial glycine-rich peptides from the hemocytes of the spider Cupiennius salei. Cellular and Molecular Life Sciences 2010, 67, 2787 -2798.
AMA StyleTommy Baumann, Urs Kämpfer, Stefan Schürch, Johann Schaller, Carlo Largiader, Wolfgang Nentwig, Lucia Kuhn-Nentwig. Ctenidins: antimicrobial glycine-rich peptides from the hemocytes of the spider Cupiennius salei. Cellular and Molecular Life Sciences. 2010; 67 (16):2787-2798.
Chicago/Turabian StyleTommy Baumann; Urs Kämpfer; Stefan Schürch; Johann Schaller; Carlo Largiader; Wolfgang Nentwig; Lucia Kuhn-Nentwig. 2010. "Ctenidins: antimicrobial glycine-rich peptides from the hemocytes of the spider Cupiennius salei." Cellular and Molecular Life Sciences 67, no. 16: 2787-2798.