Dataset Information

Identification of a precursor processing protease from the spider Cupiennius salei essential for venom neurotoxin maturation

ABSTRACT: 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 chymotrypsin – like 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.

INSTRUMENT(S): LTQ Orbitrap XL

ORGANISM(S): Cupiennius Salei (american Wandering Spider)

TISSUE(S): Venom

SUBMITTER: Nicolas Langenegger

LAB HEAD: Lucia Kuhn-Nentwig

PROVIDER: PXD007126 | Pride | 2018-01-02

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20161221_161209_B3_Chymotrypsin_NL_i01.mgf	Mgf
	20161221_161209_B3_Chymotrypsin_NL_i01.raw	Raw
	20161221_161209_B3_ProtK_NL_i01.mgf	Mgf
	20161221_161209_B3_ProtK_NL_i01.raw	Raw
	20161221_161209_B3_Trypsin_NL_i01.mgf	Mgf

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Identification of a precursor processing protease from the spider <i>Cupiennius salei</i> essential for venom neurotoxin maturation.

Langenegger Nicolas N Koua Dominique D Schürch Stefan S Heller Manfred M Nentwig Wolfgang W Kuhn-Nentwig Lucia L

The Journal of biological chemistry 20171221 6

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 10 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 neurotox ...[more]

PMID: 29269415

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