Investigation of Binding Modes and Functional Surface of Scorpion Toxins ANEP to Sodium Channels 1.7
Abstract
:1. Introduction
2. Results
2.1. Molecular Modeling of mNav1.7 (Resting and Activated States)
2.2. Molecular Docking Study
2.3. Molecular Dynamics Simulations
2.4. Construction, Expression and Purification of Mutants
2.5. Analgesic Activities of ANEP Mutants
3. Discussion
4. Conclusions
5. Materials and Methods
5.1. Homology Models
5.2. IIS1–S4 Domains Molecular Dynamics Simulations
5.3. Molecular Docking
5.4. The Complex Molecular Dynamics Simulations
5.5. Strains, Materials, and Animals
5.6. Site-Directed Mutagenesis of ANEP
5.7. Expression and Purification of ANEP and Its Mutants
5.8. Analgesic Activity Assays
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Group 1 | Dosage (mg/kg) | Number of Writhes (Mean ± SEM) | Inhibition Efficiency (%) 1 | Relative Activity (%) |
---|---|---|---|---|
NS | - | 47.67 ± 1.77 | - | - |
rANEP | 1.5 | 29.00 ± 3.41 * | 39.17% | 100 |
rANEP-N8A | 1.5 | 21.60 ± 3.7 *,2 | 54.68% | 132 |
rANEP-V12A | 1.5 | 20.33 ± 2.98 *,2 | 57.35% | 143 |
rANEP-S13A | 1.5 | 29.70 ± 2.5 * | 37.69% | 96 |
rANEP-L15A | 1.5 | 22.00 ± 1.25 *,2 | 53.85% | 132 |
rANEP-W16A | 1.5 | 23.88 ± 1.55 * | 49.90% | 121 |
rANEP-E24A | 1.5 | 20.50 ± 3.26 *,2 | 57.00% | 141 |
rANEP-Y28A | 1.5 | 24.75 ± 2.6 * | 48.06% | 114 |
rANEP-W36A | 1.5 | 13.56 ± 2.36 *,2 | 71.55% | 214 |
rANEP-T37A | 1.5 | 15.89 ± 3.11 *,2 | 66.67% | 183 |
rANEP-W38A | 1.5 | 15.56 ± 2.47 *,2 | 67.36% | 186 |
rANEP-G39A | 1.5 | 21.93 ± 2.1 *,2 | 55.12% | 132 |
rANEP-L40A | 1.5 | 30.33 ± 2.8 * | 35.71% | 93 |
rANEP-C60A | 1.5 | 35.21 ± 2.1 *,2 | 26.14% | 67 |
rANEP-K64⊿ | 1.5 | 14.98 ± 3.16 *,2 | 68.58% | 175 |
Name | Nucleotide Sequence (5′-3′) | Orientation |
---|---|---|
ANEP-F | CATGCCATGGGACATCATCATCATCATCACGATGGATATATAAGAGGAAGTAACGGATG | Sense |
ANEP-R | CGGGATCCTTACTTTTTGCCACCGCATGTATTACTTTCA | Antisense |
N8A | CATGCCATGGGACACCACCACCACCACCACGATGGATATATAAGAGGAAGTGCGGGATG | Sense |
V12A | AACGGATGCAAGGCGTCATGC | Sense |
ATGACGCTTGCATCCGTTACT | Antisense | |
S13A | CAAGGTTGCGTGCTTATGGGGA | Sense |
CATAAGCACGCAACCTTGCATCC | Antisense | |
L15A | TCATGCGCGTGGGGAAATG | Sense |
TCCCCACGCGCATGAAACCTT | Antisense | |
W16A | TCATGCTTAGCGGGAAATGAC | Sense |
GACATTTCCCGCTAAGCATGAA | Antisense | |
E24A | CAATAAAGCGTGCAGAGC | Sense |
TGCACGCTTTATTGCAACCGTCATT | Antisense | |
Y28A | AGAGCGGCGGGTGCCTCTT | Sense |
GAGGCACCCGCCGCTCTG | Antisense | |
W36A | TATGGTTATTGCGCGACCTGGGGA | Sense |
AAGTCCCCAGGTCGCGCAATAACCATAAGA | Antisense | |
T37A | TGCTGGGCGTGGGGACT | Sense |
TCCCCACGCCCAGCAATAA | Antisense | |
W38A | ACCGCGGGACTTGCATGC | Sense |
TCCCGCGGTCCAGCAATAAC | Antisense | |
G39A | ACCTGGGCGCTTGCATGCTG | sense |
CAGCATGCAAGCGCCCAGGT | Antisense | |
L40A | TGGGGAGCGGCATGCTG | Sense |
CAGCATGCCGCTCCCCA | Antisense | |
C60A | CGGGATCCTTACTTTTTGCCACCCGCTGTATTACT | Antisense |
K64⊿ | CGGGATCCTTATTTGCCACCGCATGTATTACTT | Antisense |
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Song, Y.; Liu, Z.; Zhang, Q.; Li, C.; Jin, W.; Liu, L.; Zhang, J.; Zhang, J. Investigation of Binding Modes and Functional Surface of Scorpion Toxins ANEP to Sodium Channels 1.7. Toxins 2017, 9, 387. https://doi.org/10.3390/toxins9120387
Song Y, Liu Z, Zhang Q, Li C, Jin W, Liu L, Zhang J, Zhang J. Investigation of Binding Modes and Functional Surface of Scorpion Toxins ANEP to Sodium Channels 1.7. Toxins. 2017; 9(12):387. https://doi.org/10.3390/toxins9120387
Chicago/Turabian StyleSong, Yongbo, Zeyu Liu, Qi Zhang, Chunming Li, Wei Jin, Lili Liu, Jianye Zhang, and Jinghai Zhang. 2017. "Investigation of Binding Modes and Functional Surface of Scorpion Toxins ANEP to Sodium Channels 1.7" Toxins 9, no. 12: 387. https://doi.org/10.3390/toxins9120387