Insecticidal peptides from the theraposid spider Brachypelma albiceps: An NMR-based model of Ba2

Corzo, Gerardo; Bernard, Cédric; Clement, Herlinda; Villegas, Elba; Bosmans, Frank; Tytgat, Jan; Possani, Lourival D.; Darbon, Hervé; Alagón, Alejandro

doi:10.1016/j.bbapap.2009.04.004

Cited by 32 publications

(28 citation statements)

References 36 publications

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“…GTx4 series are similar to ESTX [42], BsTx [43], JZTX-47, 48 [16], and Ba1, 2 [44] (Figure 3(a)). They are characterized as conserved six cysteine residues.…”

Section: Resultsmentioning

confidence: 99%

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Molecular Cloning and Sequence Analysis of the cDNAs Encoding Toxin-Like Peptides from the Venom Glands of Tarantula Grammostola rosea

Kimura

Ono

Kubo

2012

International Journal of Peptides

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Tarantula venom glands produce a large variety of bioactive peptides. Here we present the identification of venom components obtained by sequencing clones isolated from a cDNA library prepared from the venom glands of the Chilean common tarantula, Grammostola rosea. The cDNA sequences of about 1500 clones out of 4000 clones were analyzed after selection using several criteria. Forty-eight novel toxin-like peptides (GTx1 to GTx7, and GTx-TCTP and GTx-CRISP) were predicted from the nucleotide sequences. Among these peptides, twenty-four toxins are ICK motif peptides, eleven peptides are MIT1-like peptides, and seven are ESTX-like peptides. Peptides similar to JZTX-64, aptotoxin, CRISP, or TCTP are also obtained. GTx3 series possess a cysteine framework that is conserved among vertebrate MIT1, Bv8, prokineticins, and invertebrate astakines. GTx-CRISP is the first CRISP-like protein identified from the arthropod venom. Real-time PCR revealed that the transcripts for TCTP-like peptide are expressed in both the pereopodal muscle and the venom gland. Furthermore, a unique peptide GTx7-1, whose signal and prepro sequences are essentially identical to those of HaTx1, was obtained.

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“…GTx4 series are similar to ESTX [42], BsTx [43], JZTX-47, 48 [16], and Ba1, 2 [44] (Figure 3(a)). They are characterized as conserved six cysteine residues.…”

Section: Resultsmentioning

confidence: 99%

“…The effects of ESTX and BsTx are not clear. Ba1 and Ba2 are insecticidal peptides purified from theraphosid spider Brachypelma albiceps venom and an NMR-based 3D model of Ba2 is proposed [44]. …”

Section: Resultsmentioning

confidence: 99%

Molecular Cloning and Sequence Analysis of the cDNAs Encoding Toxin-Like Peptides from the Venom Glands of Tarantula Grammostola rosea

Kimura

Ono

Kubo

2012

International Journal of Peptides

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“…Several insecticidal peptides from Brachipelma spp. have been described; toxin Bs1 from B. smithi [23] and the peptides Ba1 and Ba2 from B. albiceps [24] are peptides which have been shown to be toxic for crickets, but not for mice. An obvious difference after alignment of the peptides toxin-like R/LFEC with toxins LpTx2, LTx1 and LTx2, can be found in the additional ten amino acids in the sequence of the latter three toxins.…”

Section: Toxin Like-peptidesmentioning

confidence: 99%

Venom components from Citharischius crawshayi spider (Family Theraphosidae): exploring transcriptome, venomics, and function

Diego‐García

Peigneur

Waelkens

et al. 2010

Cell. Mol. Life Sci.

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Despite strong efforts, knowledge about the composition of the venom of many spider species remains very limited. This work is the first report of transcriptome and venom analysis of the African spider Citharischius crawshayi. We used combined protocols of transcriptomics, venomics, and biological assays to characterize the venom and genes expressed in venom glands. A cDNA library of the venom glands was constructed and used to generate expressed sequence tags (ESTs). Sequence comparisons from 236 ESTs revealed interesting and unique sequences, corresponding to toxin-like and other components. Mass spectrometrical analysis of venom fractions showed more than 600 molecular masses, some of which showed toxic activity on crickets and modulated sodium currents in DmNa(v)1 and Na(v)1.6 channels as expressed in Xenopus oocytes. Taken together, our results may contribute to a better understanding of the cellular processes involved in the transcriptome and help us to discover new components from spider venom glands with therapeutic potential.

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“…The primary structure of Ba1 comprises 39 residues and 6 half-cystines that form three disulfide bridges. The three dimensional structure of Ba1 was solved by NMR, and it contains three antiparallel β-sheets and belongs to the DDH family of spider neurotoxins [3]. Although the cell receptor of Ba1 has not been found, it is thought that its target is the voltage-gated sodium channel (Nav) of insects because of its rapid and strong insecticidal activity, which resembles that of the known Nav neurotoxins from the venom of spiders and scorpions [4, 5].…”

Section: Introductionmentioning

confidence: 99%

A comparison between the recombinant expression and chemical synthesis of a short cysteine-rich insecticidal spider peptide

Clement¹,

Flores²,

Diego‐García³

et al. 2015

J Venom Anim Toxins Incl Trop Dis

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BackgroundThe choice between heterologous expression versus chemical synthesis for synthesizing short cysteine-rich insecticidal peptides from arthropods may impact the obtainment of yields and well-folded bioactive molecules for scientific research. Therefore, two recombinant expression systems were compared to that of chemical synthesis for producing Ba1, a cysteine-rich spider neurotoxin.MethodsThe transcription of the insecticidal neurotoxin Ba1 was obtained from a cDNA library of venom glands of the spider Brachypelma albiceps. It was cloned into the pCR®2.1-TOPO® cloning vector and then introduced in two different expression vectors, pQE40 and pET28a+. Each vector was transfected into E. coli M15 and BL21 cells, respectively, and expressed under induction with isopropyl thiogalactoside (IPTG). The chemical synthesis of Ba1 was performed in an Applied Biosystems 433A peptide synthesizer.ResultsBoth expression systems pQE40 and pET28a+ expressed the His-tagged recombinant protein products, HisrDFHRBa1 and HisrBa1, respectively, as inclusion bodies. The recombinant proteins HisrDFHRBa1 and HisrBa1 presented respective molecular masses of 28,289 and 8274.6 Da, and were not biologically active. These results suggested that both HisrDFHRBa1 and HisrBa1 were oxidized after cell extraction, and that their insecticidal activities were affected by their N-terminal pro-peptides and different disulfide bridge arrangements. The respective protein expression yields for HisrDFHRBa1 and HisrBa1 were 100 μg/L and 900 μg/L of culture medium. HisrBa1 was reduced and folded under in vitro conditions. The in vitro folding of HisrBa1 produced several isoforms, one of which, after removing its N-terminal pro-peptide by enzymatic cleavage, presented elevated insecticidal activities compared to the native Ba1. Furthermore, the His-tagged protein HisrDFHRBa1 underwent enzymatic cleavage to obtain recombinant Ba1 (rBa1). As expected, the molecular mass of rBa1 was 4406.4 Da. On the other hand, Ba1 was chemically synthesized (sBa1) with a yield of 11 mg per 0.1 mmol of amino acid assembly.ConclusionsThe two recombinant insecticidal peptides and the one synthesized chemically were as active as the native Ba1; however, toxin yields differed drastically.

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Insecticidal peptides from the theraposid spider Brachypelma albiceps: An NMR-based model of Ba2

Cited by 32 publications

References 36 publications

Molecular Cloning and Sequence Analysis of the cDNAs Encoding Toxin-Like Peptides from the Venom Glands of Tarantula Grammostola rosea

Molecular Cloning and Sequence Analysis of the cDNAs Encoding Toxin-Like Peptides from the Venom Glands of Tarantula Grammostola rosea

Venom components from Citharischius crawshayi spider (Family Theraphosidae): exploring transcriptome, venomics, and function

A comparison between the recombinant expression and chemical synthesis of a short cysteine-rich insecticidal spider peptide

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