DOE PAGES title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: The Role of Disulfide Bond Replacements in Analogues of the Tarantula Toxin ProTx-II and Their Effects on Inhibition of the Voltage-Gated Sodium Ion Channel Na v 1.7

Abstract

Spider venom toxins, such as Protoxin-II (ProTx-II), have recently received much attention as selective Nav1.7 channel blockers, with potential to be developed as leads for the treatment of chronic nocioceptive pain. ProTx-II is a 30-amino acid peptide with three disulfide bonds that has been reported to adopt a well-defined inhibitory cystine knot (ICK) scaffold structure. Potential drawbacks with such peptides include poor pharmacodynamics and potential scrambling of the disulfide bonds in vivo. In order to address these issues, in the present study we report the solid-phase synthesis of lanthionine-bridged analogues of ProTx-II, in which one of the three disulfide bridges is replaced with a thioether linkage, and evaluate the biological properties of these analogues. We have also investigated the folding and disulfide bridging patterns arising from different methods of oxidation of the linear peptide precursor. Finally, we report the X-ray crystal structure of ProTx-II to atomic resolution; to our knowledge this is the first crystal structure of an ICK spider venom peptide not bound to a substrate.

Authors:
 [1];  [2];  [2];  [3];  [3];  [4];  [5];  [5];  [5];  [6];  [3];  [4]; ORCiD logo [2]
  1. Department of Chemistry, University College London, 20 Gordon Street, London WC1H 0AJ, United Kingdom
  2. Department of Chemistry, University College London, 20 Gordon Street, London WC1H 0AJ, United Kingdom
  3. Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia 20147, United States
  4. Institute of Structural and Molecular Biology, Division of Biosciences, University College London, Gower Street, London WC1E 6BT, United Kingdom, Institute of Structural and Molecular Biology, Department of Biological Sciences, Birkbeck College, University of London, London WC1E 7HX, United Kingdom
  5. European Knowledge Centre, Eisai Limited, Mosquito Way, Hatfield, Hertfordshire AL10 9SN, United Kingdom
  6. Institute of Structural and Molecular Biology, Department of Biological Sciences, Birkbeck College, University of London, London WC1E 7HX, United Kingdom
Publication Date:
Research Org.:
Univ. College London (UCL) (United Kingdom)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
OSTI Identifier:
1395186
Alternate Identifier(s):
OSTI ID: 1508270; OSTI ID: 1508271
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Published Article
Journal Name:
Journal of the American Chemical Society
Additional Journal Information:
Journal Name: Journal of the American Chemical Society Journal Volume: 139 Journal Issue: 37; Journal ID: ISSN 0002-7863
Publisher:
American Chemical Society (ACS)
Country of Publication:
United States
Language:
English
Subject:
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY

Citation Formats

Wright, Zoë V. F., McCarthy, Stephen, Dickman, Rachael, Reyes, Francis E., Sanchez-Martinez, Silvia, Cryar, Adam, Kilford, Ian, Hall, Adrian, Takle, Andrew K., Topf, Maya, Gonen, Tamir, Thalassinos, Konstantinos, and Tabor, Alethea B. The Role of Disulfide Bond Replacements in Analogues of the Tarantula Toxin ProTx-II and Their Effects on Inhibition of the Voltage-Gated Sodium Ion Channel Na v 1.7. United States: N. p., 2017. Web. doi:10.1021/jacs.7b06506.
Wright, Zoë V. F., McCarthy, Stephen, Dickman, Rachael, Reyes, Francis E., Sanchez-Martinez, Silvia, Cryar, Adam, Kilford, Ian, Hall, Adrian, Takle, Andrew K., Topf, Maya, Gonen, Tamir, Thalassinos, Konstantinos, & Tabor, Alethea B. The Role of Disulfide Bond Replacements in Analogues of the Tarantula Toxin ProTx-II and Their Effects on Inhibition of the Voltage-Gated Sodium Ion Channel Na v 1.7. United States. https://doi.org/10.1021/jacs.7b06506
Wright, Zoë V. F., McCarthy, Stephen, Dickman, Rachael, Reyes, Francis E., Sanchez-Martinez, Silvia, Cryar, Adam, Kilford, Ian, Hall, Adrian, Takle, Andrew K., Topf, Maya, Gonen, Tamir, Thalassinos, Konstantinos, and Tabor, Alethea B. Thu . "The Role of Disulfide Bond Replacements in Analogues of the Tarantula Toxin ProTx-II and Their Effects on Inhibition of the Voltage-Gated Sodium Ion Channel Na v 1.7". United States. https://doi.org/10.1021/jacs.7b06506.
@article{osti_1395186,
title = {The Role of Disulfide Bond Replacements in Analogues of the Tarantula Toxin ProTx-II and Their Effects on Inhibition of the Voltage-Gated Sodium Ion Channel Na v 1.7},
author = {Wright, Zoë V. F. and McCarthy, Stephen and Dickman, Rachael and Reyes, Francis E. and Sanchez-Martinez, Silvia and Cryar, Adam and Kilford, Ian and Hall, Adrian and Takle, Andrew K. and Topf, Maya and Gonen, Tamir and Thalassinos, Konstantinos and Tabor, Alethea B.},
abstractNote = {Spider venom toxins, such as Protoxin-II (ProTx-II), have recently received much attention as selective Nav1.7 channel blockers, with potential to be developed as leads for the treatment of chronic nocioceptive pain. ProTx-II is a 30-amino acid peptide with three disulfide bonds that has been reported to adopt a well-defined inhibitory cystine knot (ICK) scaffold structure. Potential drawbacks with such peptides include poor pharmacodynamics and potential scrambling of the disulfide bonds in vivo. In order to address these issues, in the present study we report the solid-phase synthesis of lanthionine-bridged analogues of ProTx-II, in which one of the three disulfide bridges is replaced with a thioether linkage, and evaluate the biological properties of these analogues. We have also investigated the folding and disulfide bridging patterns arising from different methods of oxidation of the linear peptide precursor. Finally, we report the X-ray crystal structure of ProTx-II to atomic resolution; to our knowledge this is the first crystal structure of an ICK spider venom peptide not bound to a substrate.},
doi = {10.1021/jacs.7b06506},
journal = {Journal of the American Chemical Society},
number = 37,
volume = 139,
place = {United States},
year = {2017},
month = {9}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
https://doi.org/10.1021/jacs.7b06506

Citation Metrics:
Cited by: 13 works
Citation information provided by
Web of Science

Save / Share:

Works referenced in this record:

Characterization of Phosphorylated Peptides Using Traveling Wave-Based and Drift Cell Ion Mobility Mass Spectrometry
journal, January 2009

  • Thalassinos, Konstantinos; Grabenauer, Megan; Slade, Susan E.
  • Analytical Chemistry, Vol. 81, Issue 1
  • DOI: 10.1021/ac801916h

Toxins and drug discovery
journal, December 2014


Interaction of Tarantula Venom Peptide ProTx-II with Lipid Membranes Is a Prerequisite for Its Inhibition of Human Voltage-gated Sodium Channel NaV1.7
journal, August 2016

  • Henriques, Sónia Troeira; Deplazes, Evelyne; Lawrence, Nicole
  • Journal of Biological Chemistry, Vol. 291, Issue 33
  • DOI: 10.1074/jbc.M116.729095

N -Chlorosuccinimide, an Efficient Reagent for On-Resin Disulfide Formation in Solid-Phase Peptide Synthesis
journal, January 2013

  • Postma, Tobias M.; Albericio, Fernando
  • Organic Letters, Vol. 15, Issue 3
  • DOI: 10.1021/ol303428d

Synthesis and Activity of Thioether-Containing Analogues of the Complement Inhibitor Compstatin
journal, May 2011

  • Knerr, Patrick J.; Tzekou, Apostolia; Ricklin, Daniel
  • ACS Chemical Biology, Vol. 6, Issue 7
  • DOI: 10.1021/cb2000378

Optimization of oxidative folding methods for cysteine-rich peptides: a study of conotoxins containing three disulfide bridges
journal, September 2010

  • Steiner, Andrew M.; Bulaj, Grzegorz
  • Journal of Peptide Science, Vol. 17, Issue 1
  • DOI: 10.1002/psc.1283

Ziconotide: a clinical update and pharmacologic review
journal, March 2013


Ring-closing metathesis in peptides
journal, September 2016


Engineering Potent and Selective Analogues of GpTx-1, a Tarantula Venom Peptide Antagonist of the Na V 1.7 Sodium Channel
journal, February 2015

  • Murray, Justin K.; Ligutti, Joseph; Liu, Dong
  • Journal of Medicinal Chemistry, Vol. 58, Issue 5
  • DOI: 10.1021/jm501765v

An Overview of the Three Dimensional Structure ofs Short Spider Toxins
journal, January 2005


Chemical Syntheses and Biological Activities of Lactam Analogues of α-Conotoxin SI , + , #
journal, December 2000

  • Hargittai, Balazs; Solé, Núria A.; Groebe, Duncan R.
  • Journal of Medicinal Chemistry, Vol. 43, Issue 25
  • DOI: 10.1021/jm990635c

Selenoether oxytocin analogues have analgesic properties in a mouse model of chronic abdominal pain
journal, January 2014

  • de Araujo, Aline Dantas; Mobli, Mehdi; Castro, Joel
  • Nature Communications, Vol. 5, Issue 1
  • DOI: 10.1038/ncomms4165

Pathway of Oxidative Folding of a 3-Disulfide α-Lactalbumin May Resemble Either BPTI Model or Hirudin Model
journal, May 2006


Oxidation of Lanthionines Renders the Lantibiotic Nisin Inactive
journal, March 2009

  • Wilson-Stanford, Shawanda; Kalli, Anastasia; Håkansson, Kristina
  • Applied and Environmental Microbiology, Vol. 75, Issue 5
  • DOI: 10.1128/AEM.01864-08

α-Conotoxin ImI Incorporating Stable Cystathionine Bridges Maintains Full Potency and Identical Three-Dimensional Structure
journal, October 2011

  • Dekan, Zoltan; Vetter, Irina; Daly, Norelle L.
  • Journal of the American Chemical Society, Vol. 133, Issue 40
  • DOI: 10.1021/ja206408q

Distinct Disulfide Isomers of μ-Conotoxins KIIIA and KIIIB Block Voltage-Gated Sodium Channels
journal, November 2012

  • Khoo, Keith K.; Gupta, Kallol; Green, Brad R.
  • Biochemistry, Vol. 51, Issue 49
  • DOI: 10.1021/bi301256s

α-Selenoconotoxins, a New Class of Potent α7 Neuronal Nicotinic Receptor Antagonists
journal, May 2006

  • Armishaw, Christopher J.; Daly, Norelle L.; Nevin, Simon T.
  • Journal of Biological Chemistry, Vol. 281, Issue 20
  • DOI: 10.1074/jbc.M512419200

Studies Examining the Relationship between the Chemical Structure of Protoxin II and Its Activity on Voltage Gated Sodium Channels
journal, July 2014

  • Park, Jae H.; Carlin, Kevin P.; Wu, Gang
  • Journal of Medicinal Chemistry, Vol. 57, Issue 15
  • DOI: 10.1021/jm500687u

Engineering Highly Potent and Selective Microproteins against Nav1.7 Sodium Channel for Treatment of Pain
journal, July 2016

  • Shcherbatko, Anatoly; Rossi, Andrea; Foletti, Davide
  • Journal of Biological Chemistry, Vol. 291, Issue 27
  • DOI: 10.1074/jbc.M116.725978

No Gain, No Pain: Na V 1.7 as an Analgesic Target
journal, August 2014

  • King, Glenn F.; Vetter, Irina
  • ACS Chemical Neuroscience, Vol. 5, Issue 9
  • DOI: 10.1021/cn500171p

Lanthionine−Somatostatin Analogs:  Synthesis, Characterization, Biological Activity, and Enzymatic Stability Studies
journal, July 1997

  • Ösapay, George; Prokai, Laszlo; Kim, Ho-Seung
  • Journal of Medicinal Chemistry, Vol. 40, Issue 14
  • DOI: 10.1021/jm960850i

Antimicrobial Leucocin Analogues with a Disulfide Bridge Replaced by a Carbocycle or by Noncovalent Interactions of Allyl Glycine Residues
journal, November 2006

  • Derksen, Darren J.; Stymiest, Jake L.; Vederas, John C.
  • Journal of the American Chemical Society, Vol. 128, Issue 44
  • DOI: 10.1021/ja066203q

The structure of spider toxin huwentoxin-II with unique disulfide linkage: Evidence for structural evolution
journal, February 2002

  • Shu, Qin; Lu, Shan-Yun; Gu, Xiao-Cheng
  • Protein Science, Vol. 11, Issue 2
  • DOI: 10.1110/ps.30502

Travelling wave ion mobility mass spectrometry studies of protein structure: biological significance and comparison with X-ray crystallography and nuclear magnetic resonance spectroscopy measurements
journal, October 2008

  • Scarff, Charlotte A.; Thalassinos, Konstantinos; Hilton, Gillian R.
  • Rapid Communications in Mass Spectrometry, Vol. 22, Issue 20
  • DOI: 10.1002/rcm.3737

Recent advances in synthetic analogues of lantibiotics: What can we learn from these?
journal, August 2014


On the use of PyAOP, a phosphonium salt derived from HOAt, in solid-phase peptide synthesis
journal, July 1997


Discovery, Synthesis, and Structure–Activity Relationships of Conotoxins
journal, April 2014

  • Akondi, Kalyana B.; Muttenthaler, Markus; Dutertre, Sébastien
  • Chemical Reviews, Vol. 114, Issue 11
  • DOI: 10.1021/cr400401e

ProTx-II, a Selective Inhibitor of Na V 1.7 Sodium Channels, Blocks Action Potential Propagation in Nociceptors
journal, August 2008

  • Schmalhofer, William A.; Calhoun, Jeffrey; Burrows, Rachel
  • Molecular Pharmacology, Vol. 74, Issue 5
  • DOI: 10.1124/mol.108.047670

Variable Substrate Preference among Phospholipase D Toxins from Sicariid Spiders
journal, April 2015

  • Lajoie, Daniel M.; Roberts, Sue A.; Zobel-Thropp, Pamela A.
  • Journal of Biological Chemistry, Vol. 290, Issue 17
  • DOI: 10.1074/jbc.M115.636951

Diaminodiacid Bridges to Improve Folding and Tune the Bioactivity of Disulfide-Rich Peptides
journal, June 2015

  • Guo, Ye; Sun, De-Meng; Wang, Feng-Liang
  • Angewandte Chemie International Edition, Vol. 54, Issue 48
  • DOI: 10.1002/anie.201500699

Diaminodiacid-based synthesis of macrocyclic peptides using 1,2,3-triazole bridges as disulfide bond mimetics
journal, January 2017


Insensitivity to pain induced by a potent selective closed-state Nav1.7 inhibitor
journal, January 2017

  • Flinspach, M.; Xu, Q.; Piekarz, A. D.
  • Scientific Reports, Vol. 7, Issue 1
  • DOI: 10.1038/srep39662

Orthogonally Protected Lanthionines:  Synthesis and Use for the Solid-Phase Synthesis of an Analogue of Nisin Ring C
journal, March 2005

  • Bregant, Sarah; Tabor, Alethea B.
  • The Journal of Organic Chemistry, Vol. 70, Issue 7
  • DOI: 10.1021/jo048222t

High-throughput production of two disulphide-bridge toxins
journal, January 2014

  • Upert, Grégory; Mourier, Gilles; Pastor, Alexandra
  • Chem. Commun., Vol. 50, Issue 61
  • DOI: 10.1039/C4CC02679A

Synthesis of the Lantibiotic Lactocin S Using Peptide Cyclizations on Solid Phase
journal, December 2009

  • Ross, Avena C.; Liu, Hongqiang; Pattabiraman, Vijaya R.
  • Journal of the American Chemical Society, Vol. 132, Issue 2
  • DOI: 10.1021/ja9095945

Peptide therapeutics: current status and future directions
journal, January 2015


Dicarba α-Conotoxin Vc1.1 Analogues with Differential Selectivity for Nicotinic Acetylcholine and GABA B Receptors
journal, June 2013

  • van Lierop, Bianca J.; Robinson, Samuel D.; Kompella, Shiva N.
  • ACS Chemical Biology, Vol. 8, Issue 8
  • DOI: 10.1021/cb4002393

Two Tarantula Peptides Inhibit Activation of Multiple Sodium Channels
journal, December 2002

  • Middleton, Richard E.; Warren, Vivien A.; Kraus, Richard L.
  • Biochemistry, Vol. 41, Issue 50
  • DOI: 10.1021/bi026546a

N,O-bisFmoc derivatives ofN-(2-hydroxy-4-methoxybenzyl)-amino acids: Useful intermediates in peptide synthesis
journal, January 1995

  • Johnson, Tony; Quibell, Martin; Sheppard, Robert C.
  • Journal of Peptide Science, Vol. 1, Issue 1
  • DOI: 10.1002/psc.310010104

Synthesis of Oxytocin Analogues with Replacement of Sulfur by Carbon Gives Potent Antagonists with Increased Stability
journal, September 2005

  • Stymiest, Jake L.; Mitchell, Bryan F.; Wong, Susan
  • The Journal of Organic Chemistry, Vol. 70, Issue 20
  • DOI: 10.1021/jo050539l

Pharmacological characterisation of the highly NaV1.7 selective spider venom peptide Pn3a
journal, January 2017

  • Deuis, Jennifer R.; Dekan, Zoltan; Wingerd, Joshua S.
  • Scientific Reports, Vol. 7, Issue 1
  • DOI: 10.1038/srep40883

An SCN9A channelopathy causes congenital inability to experience pain
journal, December 2006

  • Cox, James J.; Reimann, Frank; Nicholas, Adeline K.
  • Nature, Vol. 444, Issue 7121
  • DOI: 10.1038/nature05413

Solving the α-Conotoxin Folding Problem: Efficient Selenium-Directed On-Resin Generation of More Potent and Stable Nicotinic Acetylcholine Receptor Antagonists
journal, February 2010

  • Muttenthaler, Markus; Nevin, Simon T.; Grishin, Anton A.
  • Journal of the American Chemical Society, Vol. 132, Issue 10
  • DOI: 10.1021/ja910602h

Microwave heating in solid-phase peptide synthesis
journal, January 2012

  • Pedersen, Søren L.; Tofteng, A. Pernille; Malik, Leila
  • Chem. Soc. Rev., Vol. 41, Issue 5
  • DOI: 10.1039/C1CS15214A

The challenge of the lantibiotics: synthetic approaches to thioether-bridged peptides
journal, January 2011

  • Tabor, Alethea B.
  • Organic & Biomolecular Chemistry, Vol. 9, Issue 22
  • DOI: 10.1039/c1ob05946g

Structure and Activity of (2,8)-Dicarba-(3,12)-cystino α-ImI, an α-Conotoxin Containing a Nonreducible Cystine Analogue
journal, January 2009

  • MacRaild, Christopher A.; Illesinghe, Jayamini; Lierop, Bianca J. van
  • Journal of Medicinal Chemistry, Vol. 52, Issue 3
  • DOI: 10.1021/jm8011504

Synthesis of Peptides Containing Overlapping Lanthionine Bridges on the Solid Phase: An Analogue of Rings D and E of the Lantibiotic Nisin
journal, August 2011

  • Mothia, Begum; Appleyard, Antony N.; Wadman, Sjoerd
  • Organic Letters, Vol. 13, Issue 16
  • DOI: 10.1021/ol201548m

Synthesis and conformational properties of the lanthionine-bridged opioid peptide [D-AlaL2,AlaL5]enkephalin as determined by NMR and computer simulations
journal, October 1992

  • Polinsky, Alexander; Cooney, Michael G.; Toy-Palmer, Anna
  • Journal of Medicinal Chemistry, Vol. 35, Issue 22
  • DOI: 10.1021/jm00100a026

The Tarantula Toxins ProTx-II and Huwentoxin-IV Differentially Interact with Human Na v 1.7 Voltage Sensors to Inhibit Channel Activation and Inactivation
journal, September 2010

  • Xiao, Yucheng; Blumenthal, Kenneth; Jackson, James O.
  • Molecular Pharmacology, Vol. 78, Issue 6
  • DOI: 10.1124/mol.110.066332

Tarantula Huwentoxin-IV Inhibits Neuronal Sodium Channels by Binding to Receptor Site 4 and Trapping the Domain II Voltage Sensor in the Closed Configuration
journal, October 2008

  • Xiao, Yucheng; Bingham, Jon-Paul; Zhu, Weiguo
  • Journal of Biological Chemistry, Vol. 283, Issue 40
  • DOI: 10.1074/jbc.M708447200

Solid Supported Chemical Syntheses of Both Components of the Lantibiotic Lacticin 3147
journal, September 2011

  • Liu, Wei; Chan, Alice S. H.; Liu, Hongqiang
  • Journal of the American Chemical Society, Vol. 133, Issue 36
  • DOI: 10.1021/ja206017p

Mutations in SCN9A, encoding a sodium channel alpha subunit, in patients with primary erythermalgia
journal, March 2004


Molecular Interactions of the Gating Modifier Toxin ProTx-II with Nav1.5
journal, April 2007

  • Smith, Jaime J.; Cummins, Theodore R.; Alphy, Sujith
  • Journal of Biological Chemistry, Vol. 282, Issue 17
  • DOI: 10.1074/jbc.M610462200

Traveling-Wave Ion Mobility Mass Spectrometry Analysis of Isomeric Modified Peptides Arising from Chemical Cross-Linking
journal, December 2010

  • Santos, Luiz F. A.; Iglesias, Amadeu H.; Pilau, Eduardo J.
  • Journal of the American Society for Mass Spectrometry, Vol. 21, Issue 12
  • DOI: 10.1016/j.jasms.2010.08.017