Pharmacological characterization of potent and selective NaV1.7 inhibitors engineered from Chilobrachys jingzhao tarantula venom peptide JzTx-V
| Autor: | John B. Jordan, Justin K. Murray, Kristin L. Andrews, Licheng Shi, Philippe Favreau, Ruoyuan Yin, Violeta Yu, Josie H. Lee, Kelvin K. C. Sham, Bin Wu, Kaustav Biswas, Dong Liu, Anruo Zou, Joseph Ligutti, Jason Long, Les P. Miranda, Bryan D. Moyer, Reto Stöcklin |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
Male
0301 basic medicine Science and Technology Workforce Patch-Clamp Techniques Physiology Drug Evaluation Preclinical Action Potentials Spider Venoms lcsh:Medicine Peptide Toxicology Pathology and Laboratory Medicine Careers in Research Protein Engineering Nerve Fibers Animal Cells Ganglia Spinal Medicine and Health Sciences Toxins Macromolecular Engineering lcsh:Science Neurons chemistry.chemical_classification Analgesics Multidisciplinary Peptide analog biology Chemistry NAV1.7 Voltage-Gated Sodium Channel Recombinant Proteins Cell biology Peptide Conformation Electrophysiology Professions Engineering and Technology Synthetic Biology Cellular Types Research Article Biotechnology Sodium Channel Blockers Cell Binding Cell Physiology Science Policy Toxic Agents Neurophysiology Bioengineering Tetrodotoxin Inhibitory postsynaptic potential Membrane Potential Cell Line Chilobrachys jingzhao Structure-Activity Relationship 03 medical and health sciences Physical Stimulation Animals Humans Patch clamp Binding site Nuclear Magnetic Resonance Biomolecular Nerve Fibers Unmyelinated Venoms Sodium channel lcsh:R Biology and Life Sciences Engineers Cell Biology biology.organism_classification Mice Inbred C57BL 030104 developmental biology Amino Acid Substitution Synthetic Bioengineering Cellular Neuroscience People and Places Mutagenesis Site-Directed Population Groupings lcsh:Q Capsaicin Peptides Neuroscience |
| Zdroj: | PLoS ONE, Vol 13, Iss 5, p e0196791 (2018) PLoS ONE |
| ISSN: | 1932-6203 |
| Popis: | Identification of voltage-gated sodium channel NaV1.7 inhibitors for chronic pain therapeutic development is an area of vigorous pursuit. In an effort to identify more potent leads compared to our previously reported GpTx-1 peptide series, electrophysiology screening of fractionated tarantula venom discovered the NaV1.7 inhibitory peptide JzTx-V from the Chinese earth tiger tarantula Chilobrachys jingzhao. The parent peptide displayed nominal selectivity over the skeletal muscle NaV1.4 channel. Attribute-based positional scan analoging identified a key Ile28Glu mutation that improved NaV1.4 selectivity over 100-fold, and further optimization yielded the potent and selective peptide leads AM-8145 and AM-0422. NMR analyses revealed that the Ile28Glu substitution changed peptide conformation, pointing to a structural rationale for the selectivity gains. AM-8145 and AM-0422 as well as GpTx-1 and HwTx-IV competed for ProTx-II binding in HEK293 cells expressing human NaV1.7, suggesting that these NaV1.7 inhibitory peptides interact with a similar binding site. AM-8145 potently blocked native tetrodotoxin-sensitive (TTX-S) channels in mouse dorsal root ganglia (DRG) neurons, exhibited 30- to 120-fold selectivity over other human TTX-S channels and exhibited over 1,000-fold selectivity over other human tetrodotoxin-resistant (TTX-R) channels. Leveraging NaV1.7-NaV1.5 chimeras containing various voltage-sensor and pore regions, AM-8145 mapped to the second voltage-sensor domain of NaV1.7. AM-0422, but not the inactive peptide analog AM-8374, dose-dependently blocked capsaicin-induced DRG neuron action potential firing using a multi-electrode array readout and mechanically-induced C-fiber spiking in a saphenous skin-nerve preparation. Collectively, AM-8145 and AM-0422 represent potent, new engineered NaV1.7 inhibitory peptides derived from the JzTx-V scaffold with improved NaV selectivity and biological activity in blocking action potential firing in both DRG neurons and C-fibers. |
| Databáze: | OpenAIRE |
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