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The structure, dynamics and selectivity profile of a NaV1.7 potency-optimised huwentoxin-IV variant.
PLoS One. 2017; 12(3):e0173551.Plos

Abstract

Venom-derived peptides have attracted much attention as potential lead molecules for pharmaceutical development. A well-known example is Huwentoxin-IV (HwTx-IV), a peptide toxin isolated from the venom of the Chinese bird-eating spider Haplopelma schmitdi. HwTx-IV was identified as a potent blocker of a human voltage-gated sodium channel (hNaV1.7), which is a genetically validated analgesic target. The peptide was promising as it showed high potency at NaV1.7 (IC50 ~26 nM) and selectivity over the cardiac NaV subtype (NaV1.5). Mutagenesis studies aimed at optimising the potency of the peptide resulted in the development of a triple-mutant of HwTx-IV (E1G, E4G, Y33W, m3-HwTx-IV) with significantly increased potency against hNaV1.7 (IC50 = 0.4 ± 0.1 nM) without increased potency against hNaV1.5. The activity of m3-HwTx-IV against other NaV subtypes was, however, not investigated. Similarly, the structure of the mutant peptide was not characterised, limiting the interpretation of the observed increase in potency. In this study we produced isotope-labelled recombinant m3-HwTx-IV in E. coli, which enabled us to characterise the atomic-resolution structure and dynamics of the peptide by NMR spectroscopy. The results show that the structure of the peptide is not perturbed by the mutations, whilst the relaxation studies reveal that residues in the active site of the peptide undergo conformational exchange. Additionally, the NaV subtype selectivity of the recombinant peptide was characterised, revealing potent inhibition of neuronal NaV subtypes 1.1, 1.2, 1.3, 1.6 and 1.7. In parallel to the in vitro studies, we investigated NaV1.7 target engagement of the peptide in vivo using a rodent pain model, where m3-HwTx-IV dose-dependently suppressed spontaneous pain induced by the NaV1.7 activator OD1. Thus, our results provide further insight into the structure and dynamics of this class of peptides that may prove useful in guiding the development of inhibitors with improved selectivity for analgesic NaV subtypes.

Authors+Show Affiliations

Centre for Advanced Imaging, The University of Queensland, St Lucia, QLD, Australia. Institute for Molecular Bioscience, The University of Queensland, St Lucia, QLD, Australia.Institute for Molecular Bioscience, The University of Queensland, St Lucia, QLD, Australia.Institute for Molecular Bioscience, The University of Queensland, St Lucia, QLD, Australia.Centre for Advanced Imaging, The University of Queensland, St Lucia, QLD, Australia.Institute for Molecular Bioscience, The University of Queensland, St Lucia, QLD, Australia.Institute for Molecular Bioscience, The University of Queensland, St Lucia, QLD, Australia. School of Biomedical Sciences, The University of Queensland, St Lucia, QLD, Australia.Institute for Molecular Bioscience, The University of Queensland, St Lucia, QLD, Australia.Institute for Molecular Bioscience, The University of Queensland, St Lucia, QLD, Australia.Centre for Advanced Imaging, The University of Queensland, St Lucia, QLD, Australia.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

28301520

Citation

Rahnama, Sassan, et al. "The Structure, Dynamics and Selectivity Profile of a NaV1.7 Potency-optimised huwentoxin-IV Variant." PloS One, vol. 12, no. 3, 2017, pp. e0173551.
Rahnama S, Deuis JR, Cardoso FC, et al. The structure, dynamics and selectivity profile of a NaV1.7 potency-optimised huwentoxin-IV variant. PLoS ONE. 2017;12(3):e0173551.
Rahnama, S., Deuis, J. R., Cardoso, F. C., Ramanujam, V., Lewis, R. J., Rash, L. D., King, G. F., Vetter, I., & Mobli, M. (2017). The structure, dynamics and selectivity profile of a NaV1.7 potency-optimised huwentoxin-IV variant. PloS One, 12(3), e0173551. https://doi.org/10.1371/journal.pone.0173551
Rahnama S, et al. The Structure, Dynamics and Selectivity Profile of a NaV1.7 Potency-optimised huwentoxin-IV Variant. PLoS ONE. 2017;12(3):e0173551. PubMed PMID: 28301520.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The structure, dynamics and selectivity profile of a NaV1.7 potency-optimised huwentoxin-IV variant. AU - Rahnama,Sassan, AU - Deuis,Jennifer R, AU - Cardoso,Fernanda C, AU - Ramanujam,Venkatraman, AU - Lewis,Richard J, AU - Rash,Lachlan D, AU - King,Glenn F, AU - Vetter,Irina, AU - Mobli,Mehdi, Y1 - 2017/03/16/ PY - 2016/11/02/received PY - 2017/02/23/accepted PY - 2017/3/17/entrez PY - 2017/3/17/pubmed PY - 2017/9/8/medline SP - e0173551 EP - e0173551 JF - PloS one JO - PLoS ONE VL - 12 IS - 3 N2 - Venom-derived peptides have attracted much attention as potential lead molecules for pharmaceutical development. A well-known example is Huwentoxin-IV (HwTx-IV), a peptide toxin isolated from the venom of the Chinese bird-eating spider Haplopelma schmitdi. HwTx-IV was identified as a potent blocker of a human voltage-gated sodium channel (hNaV1.7), which is a genetically validated analgesic target. The peptide was promising as it showed high potency at NaV1.7 (IC50 ~26 nM) and selectivity over the cardiac NaV subtype (NaV1.5). Mutagenesis studies aimed at optimising the potency of the peptide resulted in the development of a triple-mutant of HwTx-IV (E1G, E4G, Y33W, m3-HwTx-IV) with significantly increased potency against hNaV1.7 (IC50 = 0.4 ± 0.1 nM) without increased potency against hNaV1.5. The activity of m3-HwTx-IV against other NaV subtypes was, however, not investigated. Similarly, the structure of the mutant peptide was not characterised, limiting the interpretation of the observed increase in potency. In this study we produced isotope-labelled recombinant m3-HwTx-IV in E. coli, which enabled us to characterise the atomic-resolution structure and dynamics of the peptide by NMR spectroscopy. The results show that the structure of the peptide is not perturbed by the mutations, whilst the relaxation studies reveal that residues in the active site of the peptide undergo conformational exchange. Additionally, the NaV subtype selectivity of the recombinant peptide was characterised, revealing potent inhibition of neuronal NaV subtypes 1.1, 1.2, 1.3, 1.6 and 1.7. In parallel to the in vitro studies, we investigated NaV1.7 target engagement of the peptide in vivo using a rodent pain model, where m3-HwTx-IV dose-dependently suppressed spontaneous pain induced by the NaV1.7 activator OD1. Thus, our results provide further insight into the structure and dynamics of this class of peptides that may prove useful in guiding the development of inhibitors with improved selectivity for analgesic NaV subtypes. SN - 1932-6203 UR - https://www.unboundmedicine.com/medline/citation/28301520/The_structure_dynamics_and_selectivity_profile_of_a_NaV1_7_potency_optimised_huwentoxin_IV_variant_ L2 - http://dx.plos.org/10.1371/journal.pone.0173551 DB - PRIME DP - Unbound Medicine ER -