Dataset Information

Cupiennius salei Hyal LC-MSMS - Isolation, N-glycosylations and Function of a Hyaluronidase-Like Enzyme from the Venom of the Spider Cupiennius salei

ABSTRACT: Hyaluronidases are important venom components by acting as spreading factor of neurotoxins. In several studies this spreading effect was tested on vertebrate tissue. However, data about the spreading activity on invertebrates, which are the main prey organisms of spiders, are lacking. In this study, a hyaluronidase-like enzyme was isolated from the venom of the spider Cupiennius salei. The amino acid sequence of the enzyme was solved by cDNA analysis of the venom gland transcriptome and confirmed by protein analytics. Two complex N-linked glycans, which akin honey bee hyaluronidase glycosylations, were identified by tandem mass spectrometry.

INSTRUMENT(S): Q Exactive

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

TISSUE(S): Venom Apparatus

SUBMITTER: Christian Trachsel

LAB HEAD: Lucia Kuhn-Nentwig

PROVIDER: PXD002489 | Pride | 2015-12-10

REPOSITORIES: Pride

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Publications

A Randomized Double Blind Clinical Trial on a Sabgh Formulation for Patients With Vitiligo.

Hosseinkhani Ayda A Montaseri Hashem H Sodaifi Manouchehr M Mohagheghzadeh Abdolali A Seradj Hassan H Afshari-Jouybari Hassan H

Journal of evidence-based complementary & alternative medicine 20150407 4

<h4>Background</h4>The cosmetic problem that vitiligo produces affects patients psychologically. Many patients with vitiligo are suggested to cover their white skin patches with cosmetic products. There are formulations in traditional Iranian pharmacy to color these white skin patches. In this study, one of these formulations was compared with a cosmetic formulation.<h4>Methods</h4>Two groups of patients were selected. One group used a marketed formulation and other group used a traditional Iran ...[more]

PMID: 25851025

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Project description:As biopharmaceuticals, recombinant proteins have become indispensable tools in medicine. An increasing demand, not only in quantity but also in diversity, drives the constant development and improvement of production platforms. The N-glycosylation pattern on biopharmaceuticals plays important roles in activity, serum half-life and immunogenicity. Therefore, production platforms with tailored protein N-glycosylation are of great interest. Plant-based systems have already demonstrated their potential to produce pharmaceutically relevant recombinant proteins, although their N-glycan patterns differ from those in humans. Plants have shown great plasticity towards the manipulation of their glycosylation machinery, and some have already been glyco-engineered in order to avoid the attachment of plant-typical, putatively immunogenic sugar residues. This resulted in complex-type N-glycans with a core structure identical to the human one. Compared to humans, plants lack the ability to elongate these N-glycans with β1,4-linked galactoses and terminal sialic acids. However, these modifications, which require the activity of several mammalian enzymes, have already been achieved for Nicotiana benthamiana and the moss Physcomitrella. Here, we present the first step towards sialylation of recombinant glycoproteins in Physcomitrella, human β1,4-linked terminal N-glycan galactosylation, which was achieved by the introduction of a chimeric β1,4-galactosyltransferase (FTGT). This chimeric enzyme consists of the moss α1,4-fucosyltransferase transmembrane domain, fused to the catalytic domain of the human β1,4-galactosyltransferase. Stable FTGT expression led to the desired β1,4-galactosylation. However, additional pentoses of unknown identity were also observed. The nature of these pentoses was subsequently determined by Western blot and enzymatic digestion followed by mass spectrometric analysis and resulted in their identification as α-linked arabinoses. Since a pentosylation of β1,4-galactosylated N-glycans was reported earlier, e.g. on recombinant human erythropoietin produced in glyco-engineered Nicotiana tabacum, this phenomenon is of a more general importance for plant-based production platforms. Arabinoses, which are absent in humans, may prevent the full humanization of plant-derived products. Therefore, the identification of these pentoses as arabinoses is important as it creates the basis for their abolishment to ensure the production of safe biopharmaceuticals in plant-based systems.

Project description:Assassin bugs (Hemiptera: Heteroptera: Reduviidae) are venomous insects that prey on invertebrates. Assassin bug venom has features in common with venoms from other animals, such as paralysing and lethal activity when injected, and a molecular composition that includes disulfide-rich peptide neurotoxins. Uniquely, this venom also has strong liquefying activity that has been hypothesised to facilitate feeding through the narrow channel of the proboscis—a structure inherited from sap- and phloem-feeding phytophagous hemipterans and adapted during the evolution of Heteroptera into a fang and feeding structure. However, further understanding of the function of assassin bug venom is impeded by the lack of proteomic studies detailing its molecular composition. In addition, the lack of knowledge regarding venoms of predaceous reduviids limits our understanding of how the venoms of the blood-feeding kissing bugs (Reduviidae: Triatominae) evolved to facilitate hematophagy. By using a combined transcriptomic/proteomic approach we show that the venom proteome of the harpactorine assassin bug Pristhesancus plagipennis includes a complex suite of >100 proteins comprising disulfide-rich peptides, CUB-domain proteins, cystatins, putative cytolytic toxins, triabin-like protein, odorant binding protein, serine proteases, catabolic enzymes, putative nutrient-binding proteins, plus eight families of proteins without homology to characterised proteins. Serine proteases, CUB domain proteins and other novel proteins in the 10–16 kDa mass range, as well as putative cytolytic toxins, were the most abundant venom components. Thus, in addition to putative neurotoxins, assassin bug venom includes a high proportion of enzymatic and cytolytic venom components well suited to tissue liquefaction. While some protein families such as lipocalin/triabins occur in the venoms of both predaceous and blood-feeding reduviids, the composition of venoms in these two groups differs markedly. These results provide insights into the venom evolution in the insect suborder Heteroptera.

Dataset Information

Cupiennius salei Hyal LC-MSMS - Isolation, N-glycosylations and Function of a Hyaluronidase-Like Enzyme from the Venom of the Spider Cupiennius salei

Publications

A Randomized Double Blind Clinical Trial on a Sabgh Formulation for Patients With Vitiligo.

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