Langenegger, Nicolas; Koua, Dominique; Schürch, Stefan; Heller, Manfred; Nentwig, Wolfgang; Kuhn-Nentwig, Lucia (2018). Identification of a precursor processing protease from the spider Cupiennius salei essential for venom neurotoxin maturation. Journal of biological chemistry, 293(6), pp. 2079-2090. American Society for Biochemistry and Molecular Biology 10.1074/jbc.M117.810911
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J. Biol. Chem.-2018-Langenegger-2079-90.pdf - Published Version Restricted to registered users only Available under License Publisher holds Copyright. Download (2MB) |
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.
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