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Found on all continents except Antarctica, they range in size from minute to ~1 cm in adult body length. Spiders of the family Pholcidae are interesting animals in their own right. We are not aware of any scientific work to date that has attempted to characterize venom composition of any pholcid species, and given their evolutionary depth and diversity, they provide remarkable opportunity to discover new toxins and lend insight into the broader patterns of venom diversity within spider venoms, while directly investigating a widespread urban myth. Reports of high mammalian potency emerge from rumor and non-scientific anecdote. Pholcids, in reality, are a family of spiders that has been diversifying for over 200 million years ( Dimitrov et al., 2012 Eberle et al., 2018) and are among the top six percent of the most diverse spider families (1,714 species in 94 genera, World Spider Catalog, February 2019). Commonly called “cellar spiders” some pholcids are also referred to as “daddy long-legs” and have been maligned in popular culture as having venoms with the highest mammalian potency of all spiders, that would be lethal if they only had fangs that could penetrate human skin. Pholcid spiders (Araneae: Pholcidae) are an example of disconnect between public perception, and scientific understanding. Together, the evidence we present indicates pholcid venoms are a source of interesting new peptides and proteins, and effects of bites on humans and other mammals are inconsequential. Documented bites on humans make clear that while these spiders can bite, the typical result is a mild sting with no long-lasting effects. However, crickets exhibit flaccid paralysis and regions of darkening that are not observed after P. Bioassays of crude venoms on crickets resulted in an effective paralytic dose of 3.9 μg/g, which is comparable to that of crude venoms of Plectreurys tristis and other Synspermiata taxa. Comparative evidence suggests that the neprilysin gene family is expressed in venoms across a range of spider taxa, but has undergone an expansion in the venoms of pholcids and may play a central functional role in these spiders. Neprilysins (M13 peptidases), and astacins (M12 peptidases) are the most abundant venom proteins, respectively representing 15 and 11% of the individual proteins and 32 and 20% of the tryptic peptides detected in crude venom. We identify 17 new groups of peptides (U 1−17-PHTX) most of which are homologs of known venom peptides and are predicted to have an inhibitor cysteine knot fold of these, 13 are confirmed in the proteome. The majority of the venom (51% of tryptic polypeptides and 62% of unique tryptic peptides) consists of proteins homologous to known venom toxins including enzymes (astacin metalloproteases, serine proteases and metalloendopeptidases, particularly neprilysins) and venom peptide neurotoxins.
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We also analyze the venom's potency on insects, and assemble available evidence regarding mammalian toxicity. Here we detail the venom composition of male Physocyclus mexicanus using proteomic analyses and venom-gland transcriptomes (“venomics”). Pholcid spiders (Araneae: Pholcidae), officially “cellar spiders” but popularly known as “daddy long-legs,” are renown for the potential of deadly toxic venom, even though venom composition and potency has never formally been studied. 4Arizona Proteomics Consortium, University of Arizona, Tucson, AZ, United States.3Center for Genome Research and Biocomputing, Oregon State University, Corvallis, OR, United States.2Spider Pharm, Yarnell, AZ, United States.1Department of Biology, Lewis & Clark College, Portland, OR, United States.Zobel-Thropp 1 Jennifer Mullins 1 Charles Kristensen 2 Brent A.