Solution Structures of Two Homologous Venom Peptides from Sicarius dolichocephalus

Loening, Nikolaus M.; Wilson, Zachary N; Zobel-Thropp, Pamela A.; Binford, Greta J.

doi:10.1371/journal.pone.0054401

Cited by 6 publications

(9 citation statements)

References 35 publications

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“…We have identified several ICK venom gland transcripts in all of our venom gland Haplogyne transcriptomes that are predicted to be knottins; in addition to the sicariid libraries mentioned above, we have identified several in pseudoscorpion ( Synsphronus apimelus ), archaeid ( A. mainae ), pholcid ( P. mexicanus ) and plectreurid ( P. tristis ) cDNA libraries (Binford et al , unpublished). Loening et al 53 recently solved the first NMR structures of two ICK peptides from S. dolichocephalus . The clones from U 6 -SYTX share up to 67% amino acid identity with one of these.…”

Section: Resultsmentioning

confidence: 99%

Spit and Venom from Scytodes Spiders: A Diverse and Distinct Cocktail

et al. 2013

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Spiders from the family Scytodidae have a unique prey capturing technique: they spit a zig-zagged silken glue to tether prey to a surface. Effectiveness of this sticky mixture is based on a combination of contraction and adhesion, trapping prey until the spider immobilizes it by envenomation and then feeds. We identify components expressed in Scytodes thoracica venom glands using combined transcriptomic and proteomic analyses. These include homologues of toxic proteins astacin metalloproteases and potentially toxic proteins including venom allergen, longistatin, and translationally controlled tumor protein (TCTP). We classify 19 distinct groups of candidate peptide toxins; 13 of these were detected in the venom, making up 35% of the proteome. Six have significant similarity to toxins from spider species spanning mygalomorph and nonhaplogyne araneomorph lineages, suggesting their expression in venom is phylogenetically widespread. Twelve peptide toxin groups have homologues in venom gland transcriptomes of other haplogynes. Of the transcripts, approximately 50% encode glycine-rich peptides that may contribute to sticky fibers in Scytodes spit. Fifty-one percent of the identified venom proteome is a family of proteins that is homologous to sequences from Drosophila sp. and Latrodectus hesperus with uncharacterized function. Characterization of these components holds promise for discovering new functional activity.

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Section: Resultsmentioning

confidence: 99%

Spit and Venom from Scytodes Spiders: A Diverse and Distinct Cocktail

et al. 2013

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“…There was one significant match for U 5 -Sth1a from another species; the peptide U 1 -sicaritoxin-Sdo1a [ 26 ] from the venom of Sicarius dolichocephalous has an unknown molecular function and matched with an expect value of 2×10 −8 . Potentially, this match may stem from the evolutionary relationship between Sicarius dolichocephalous and Scytodes thoracica as it has been proposed that their families (Sicariidae and Scytodidae, respectively) are sister taxa [ 39 ] that diverged at least 100 million years ago [ 40 ].…”

Section: Resultsmentioning

confidence: 99%

“…The expression of U 5 -Sth1a followed a procedure that we previously described [ 26 ]. Briefly, the sample was prepared as follows: pLICC vector containing the U 5 -Sth1a insert was used to transform E .…”

Section: Methodsmentioning

confidence: 99%

Characterization of Three Venom Peptides from the Spitting Spider Scytodes thoracica

Ariki¹,

Muñoz²,

Armitage³

et al. 2016

PLoS ONE

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We present the solution-state NMR structures and preliminary functional characterizations of three venom peptides identified from the spitting spider Scytodes thoracica. Despite little sequence identity to other venom peptides, structural characterization reveals that these peptides contain an inhibitor cystine knot motif common to many venom peptides. These are the first structures for any peptide or protein from spiders of the Scytodidae family. Many venom peptides target neuronal ion channels or receptors. However, we have not been able to determine the target of these Scytodes peptides so we can only state with certainty the channels and receptors that they do not target.

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“…Here, we assessed three commercial cell-free synthesis platforms for their ability to produce the knottin U 2 -sicaritoxin-Sdo1a (USCTX) from the six-eyed sand spider Hexophtalma dolichocephala as a model arthropod venom component. This toxin was identified by transcriptome-guided venomics and was structurally characterized by NMR spectroscopy following its expression in heterologous cells [ 21 ]. USCTX contains an inhibitory cysteine knot (ICK) motif with complex disulfide bridges, which is an added challenge for in vitro expression [ 21 ].…”

Section: Introductionmentioning

confidence: 99%

“…This toxin was identified by transcriptome-guided venomics and was structurally characterized by NMR spectroscopy following its expression in heterologous cells [ 21 ]. USCTX contains an inhibitory cysteine knot (ICK) motif with complex disulfide bridges, which is an added challenge for in vitro expression [ 21 ]. The recombinant USCTX produced by cell-free synthesis was also analyzed to determine its bioactivity.…”

Section: Introductionmentioning

confidence: 99%

A Spider Toxin Exemplifies the Promises and Pitfalls of Cell-Free Protein Production for Venom Biodiscovery

Lüddecke

Paas

Talmann

et al. 2021

Toxins

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Arthropod venoms offer a promising resource for the discovery of novel bioactive peptides and proteins, but the limited size of most species translates into minuscule venom yields. Bioactivity studies based on traditional fractionation are therefore challenging, so alternative strategies are needed. Cell-free synthesis based on synthetic gene fragments is one of the most promising emerging technologies, theoretically allowing the rapid, laboratory-scale production of specific venom components, but this approach has yet to be applied in venom biodiscovery. Here, we tested the ability of three commercially available cell-free protein expression systems to produce venom components from small arthropods, using U2-sicaritoxin-Sdo1a from the six-eyed sand spider Hexophtalma dolichocephala as a case study. We found that only one of the systems was able to produce an active product in low amounts, as demonstrated by SDS-PAGE, mass spectrometry, and bioactivity screening on murine neuroblasts. We discuss our findings in relation to the promises and limitations of cell-free synthesis for venom biodiscovery programs in smaller invertebrates.

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Solution Structures of Two Homologous Venom Peptides from Sicarius dolichocephalus

Cited by 6 publications

References 35 publications

Spit and Venom from Scytodes Spiders: A Diverse and Distinct Cocktail

Spit and Venom from Scytodes Spiders: A Diverse and Distinct Cocktail

Characterization of Three Venom Peptides from the Spitting Spider Scytodes thoracica

A Spider Toxin Exemplifies the Promises and Pitfalls of Cell-Free Protein Production for Venom Biodiscovery

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