µ-Conotoxins Targeting the Human Voltage-Gated Sodium Channel Subtype NaV1.7
Abstract
:1. Introduction
2. Results
2.1. Oxidation of Native µ-Conotoxins and Analogues
2.2. Selectivity of µ-Conotoxins for hNaV Channel Subtypes
2.3. A Limited Number of µ-Conotoxins Display Inhibitory Potency at hNaV1.7
2.4. Charged Residues in Loop 3 Affect Selectivity for hNaV1.7
2.5. Double and Triple SxIIIC Mutants
2.6. Key Structural Differences between hNaV1.7 Inhibiting µ-Conotoxins
2.7. N-Terminal Deletion Does Not Significantly Affect the Potency of SxIIIC
2.8. Loop 1 Truncation Reduces Potency of SxIIIC at hNaV1.7
3. Discussion
4. Methods
4.1. Peptide Synthesis
4.2. Oxidation
4.3. Cell Culture
4.4. Whole-Cell Patch-Clamp Electrophysiology Assays
5. Data Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Subtype | SxIIIC a | SmIIIA | KIIIA |
---|---|---|---|
NaV1.1 | 132 ± 12 | 235 ± 15.0 | 136 ± 18 |
NaV1.2 | 364 ± 54 | 172 ± 49 | 186 ± 32 |
NaV1.3 | 89 ± 11 | 95 ± 9 | >1000 |
NaV1.4 | 15 ± 11 | 14 ± 2 | 67 ± 15 |
NaV1.5 | >1000 | >1000 | >1000 |
NaV1.6 | 125 ± 11 | 106 ± 37 | 762 ± 77 |
NaV1.7 | 152 ± 22 | 41 ± 4 | 379 ± 43 |
NaV1.8 | >1000 | >1000 | >1000 |
µ-Conotoxin | Sequence | Inhibitory Potency at hNaV1.7 |
---|---|---|
GIIIA | RDCCTOOKKCKDRQCKOQ-RCCA * | No |
GIIIB | RDCCTOORKCKDRRCKOM-KCCA * | No |
GIIIC | RDCCTOOKKCKDRRCKOL-KCCA * | No |
TIIIA | RHGCCKGOKGCSSRECROQ-HCC * | No |
CnIIIC | GCCNGPKGCSSKWCRDHARCC * | Yes |
KIIIA | CCN—CSSKWCRDHSRCC * | Yes |
SIIIA | ZNCCNG—GCSSKWCRDHARCC * | No |
SmIIIA | ZRCCNGRRGCSSRWCRDHSRCC * | Yes |
SxIIIC | RGCCNGRGGCSSRWCRDHARCC * | Yes |
Analogue | hNaV1.7 | Fold Difference to SxIIIC | hNav1.4 | Fold Difference to SxIIIC |
---|---|---|---|---|
SxIIIC | 152.2 ± 26.8 | − | 12.8 ± 13.0 | − |
[Δ1,2]SxIIIC | 106.2 ± 19.4 | ns | 96.3 ± 13.0 | ↓7-fold |
[Δ7,8]SxIIIC | 313.6 ± 36.6 | ↓2-fold | − | − |
[Δ6–9]SxIIIC | 317.9 ± 100.0 | ↓2-fold | − | − |
[G8R]SxIIIC | 155.7 ± 18.9 | ns | − | − |
[Loop1R]KIIIA | 305.1 ± 23.9 | ↓2-fold * | − | − |
[R16A]SxIIIC | 334.1 ± 25.5 | ↓2-fold | 38.0 ± 13.0 | ↓3-fold |
[R16H]SxIIIC | 66.5 ± 2.5 | ↑2-fold | 80.3 ± 24.7 | ↓6-fold |
[R16Q]SxIIIC | 151.3 ± 24.0 | ns | 99.5 ± 12.9 | ↓7-fold |
[D17A]SxIIIC | 123.5 ± 18.1 | ns | 14.6 ± 7.7 | ns |
[R20A]SxIIIC | 142.3 ± 12.9 | ns | 64.8 ± 15.4 | ↓5-fold |
[Δ1,2; R16H] | 666.6 ± 211.6 | ↓4-fold | 221.4 ± 26.9 | ↓17-fold |
[Δ1,2; R20A] | 481.8 ± 646.9 | ↓3-fold | 222.3 ± 70.4 | ↓17-fold |
[R16H; R20A] | 888.5 ± 916.4 | ↓6-fold | 247.8 ± 57.7 | ↓19-fold |
[Δ1,2; R16H; R20A] | >1000 | − | >1000 | − |
µ-Conotoxin | Selectivity Profile |
---|---|
SxIIIC | 1.4 > 1.3 > 1.6 ≈ 1.1 ≈ 1.7 > 1.2 >> 1.5 ≈ 1.8 |
SmIIIA | 1.4 > 1.7 > 1.3 ≈ 1.6 > 1.2 > 1.1 >> 1.5 ≈ 1.8 |
KIIIA | 1.4 > 1.1 > 1.2 > 1.7 > 1.6 > 1.3 >> 1.5 ≈ 1.8 |
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McMahon, K.L.; Tran, H.N.T.; Deuis, J.R.; Craik, D.J.; Vetter, I.; Schroeder, C.I. µ-Conotoxins Targeting the Human Voltage-Gated Sodium Channel Subtype NaV1.7. Toxins 2022, 14, 600. https://doi.org/10.3390/toxins14090600
McMahon KL, Tran HNT, Deuis JR, Craik DJ, Vetter I, Schroeder CI. µ-Conotoxins Targeting the Human Voltage-Gated Sodium Channel Subtype NaV1.7. Toxins. 2022; 14(9):600. https://doi.org/10.3390/toxins14090600
Chicago/Turabian StyleMcMahon, Kirsten L., Hue N. T. Tran, Jennifer R. Deuis, David J. Craik, Irina Vetter, and Christina I. Schroeder. 2022. "µ-Conotoxins Targeting the Human Voltage-Gated Sodium Channel Subtype NaV1.7" Toxins 14, no. 9: 600. https://doi.org/10.3390/toxins14090600