Ancestral protein resurrection and engineering opportunities of the mamba aminergic toxins

Blanchet, Guillaume; Alili, Doria; Protte, Adèle; Upert, Grégory; Gilles, Nicolas; Tepshi, Livia; Stura, E.A.; Mourier, Gilles; Servent, Denis

doi:10.1038/s41598-017-02953-0

Cited by 16 publications

(13 citation statements)

References 47 publications

(63 reference statements)

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“…Already, 3FTXs have proven to be an efficient tool to analyze various receptor types, and to study diseases like Parkinson’s disease, myasthenia gravis and cancer [ 64 , 65 , 66 , 67 , 68 , 69 , 70 , 71 ]. The aminergic toxins from mamba venom served as good candidates for protein resurrection methodology [ 72 ].…”

Section: Discussionmentioning

confidence: 99%

Proteomic Investigations of Two Pakistani Naja Snake Venoms Species Unravel the Venom Complexity, Posttranslational Modifications, and Presence of Extracellular Vesicles

Manuwar

Dreyer

Böhmert

et al. 2020

Toxins

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Latest advancement of omics technologies allows in-depth characterization of venom compositions. In the present work we present a proteomic study of two snake venoms of the genus Naja i.e., Naja naja (black cobra) and Naja oxiana (brown cobra) of Pakistani origin. The present study has shown that these snake venoms consist of a highly diversified proteome. Furthermore, the data also revealed variation among closely related species. High throughput mass spectrometric analysis of the venom proteome allowed to identify for the N. naja venom 34 protein families and for the N. oxiana 24 protein families. The comparative evaluation of the two venoms showed that N. naja consists of a more complex venom proteome than N. oxiana venom. Analysis also showed N-terminal acetylation (N-ace) of a few proteins in both venoms. To the best of our knowledge, this is the first study revealing this posttranslational modification in snake venom. N-ace can shed light on the mechanism of regulation of venom proteins inside the venom gland. Furthermore, our data showed the presence of other body proteins, e.g., ankyrin repeats, leucine repeats, zinc finger, cobra serum albumin, transferrin, insulin, deoxyribonuclease-2-alpha, and other regulatory proteins in these venoms. Interestingly, our data identified Ras-GTpase type of proteins, which indicate the presence of extracellular vesicles in the venom. The data can support the production of distinct and specific anti-venoms and also allow a better understanding of the envenomation and mechanism of distribution of toxins. Data are available via ProteomeXchange with identifier PXD018726.

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Section: Discussionmentioning

confidence: 99%

Proteomic Investigations of Two Pakistani Naja Snake Venoms Species Unravel the Venom Complexity, Posttranslational Modifications, and Presence of Extracellular Vesicles

Manuwar

Dreyer

Böhmert

et al. 2020

Toxins

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“…The snake venom TFPs greatly differ in their activity: short- and long-chain α-neurotoxins, as well as certain non-conventional neurotoxins are blocking nicotinic acetylcholine receptors (nAChRs) (Tsetlin, 2015), while another group of TFPs is attacking muscarinic acetylcholine receptors (Karlsson et al, 2000). There are toxins inhibiting β-adrenergic and other G-protein coupled receptors (GPCRs) (Blanchet et al, 2017). Among the recently found snake venom neurotoxins are mambalgins blocking the acid-sensitive ion channels (ASICs) (Brzezicki and Zakowicz, 2018).…”

Section: Introductionmentioning

confidence: 99%

From Synthetic Fragments of Endogenous Three-Finger Proteins to Potential Drugs

et al. 2019

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The proteins of the Ly6 family have a three-finger folding as snake venom α-neurotoxins, targeting nicotinic acetylcholine receptors (nAChRs), and some of them, like mammalian secreted Ly6/uPAR protein (SLURP1) and membrane-attached Ly-6/neurotoxin (Lynx1), also interact with distinct nAChR subtypes. We believed that synthetic fragments of these endogenous proteins might open new ways for drug design because nAChRs are well-known targets for developing analgesics and drugs against neurodegenerative diseases. Since interaction with nAChRs was earlier shown for synthetic fragments of the α-neurotoxin central loop II, we synthesized a 15-membered fragment of human Lynx1, its form with two Cys residues added at the N- and C-termini and forming a disulfide, as well as similar forms of human SLURP1, SLURP2, and of Drosophila sleepless protein (SSS). The IC 50 values measured in competition with radioiodinated α-bungarotoxin for binding to the membrane-bound Torpedo californica nAChR were 4.9 and 7.4 µM for Lynx1 and SSS fragments, but over 300 µM for SLURP1 or SLURP2 fragments. The affinity of these compounds for the α7 nAChR in the rat pituitary tumor-derived cell line GH4C1 was different: 13.1 and 147 µM for SSS and Lynx1 fragments, respectively. In competition for the ligand-binding domain of the α9 nAChR subunit, SSS and Lynx1 fragments had IC 50 values of about 40 µM, which correlates with the value found for the latter with the rat α9α10 nAChR expressed in the Xenopus oocytes. Thus, the activity of these synthetic peptides against muscle-type and α9α10 nAChRs indicates that they may be useful in design of novel myorelaxants and analgesics.

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“…Evolution has long been ‘experimenting’ with amino acids substitutions in proteins and peptides in order to increase venom efficiency and efficacy as well as avoid prey evasive strategies [ 27 , 28 ]. Not only that, but also synergism aroused by the presence of several isoforms of a given molecule also increases toxicity [ 29 ].…”

Section: Discussionmentioning

confidence: 99%

Crotalus durissus terrificus crotapotin naturally displays preferred positions for amino acid substitutions

Oliveira

Ferreira

Barraviera

et al. 2017

J Venom Anim Toxins Incl Trop Dis

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BackgroundClassically, Crotalus durissus terrificus (Cdt) venom can be described, according to chromatographic criteria, as a simple venom, composed of four major toxins, namely: gyroxin, crotamine, crotoxin and convulxin. Crotoxin is a non-covalent heterodimeric neurotoxin constituted of two subunits: an active phospholipase A2 and a chaperone protein, termed crotapotin. This molecule is composed of three peptide chains connected by seven disulfide bridges. Naturally occurring variants/isoforms of either crotoxin or crotapotin itself have already been reported.MethodsThe crude Cdt venom was separated by using RP-HPLC and the toxins were identified by mass spectrometry (MS). Crotapotin was purified, reduced and alkylated in order to separate the peptide chains that were further analyzed by mass spectrometry and de novo peptide sequencing.ResultsThe RP-HPLC profile of the isolated crotapotin chains already indicated that the α chain would present isoforms, which was corroborated by the MS and tandem mass spectrometry analyses.ConclusionIt was possible to observe that the Cdt crotapotin displays a preferred amino acid substitution pattern present in the α chain, at positions 31 and 40. Moreover, substitutions could also be observed in β and γ chains (one for each). The combinations of these four different peptides, with the already described chains, would produce ten different crotapotins, which is compatible to our previous observations for the Cdt venom.

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Ancestral protein resurrection and engineering opportunities of the mamba aminergic toxins

Cited by 16 publications

References 47 publications

Proteomic Investigations of Two Pakistani Naja Snake Venoms Species Unravel the Venom Complexity, Posttranslational Modifications, and Presence of Extracellular Vesicles

Proteomic Investigations of Two Pakistani Naja Snake Venoms Species Unravel the Venom Complexity, Posttranslational Modifications, and Presence of Extracellular Vesicles

From Synthetic Fragments of Endogenous Three-Finger Proteins to Potential Drugs

Crotalus durissus terrificus crotapotin naturally displays preferred positions for amino acid substitutions

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