Engineering Gain-of-Function Analogues of the Spider Venom Peptide HNTX-I, A Potent Blocker of the hNaV1.7 Sodium Channel

Yunxiao Zhang; Qiuchu Yang; Qingfeng Zhang; Dezheng Peng; Minzhi Chen; Songping Liang; Xi Zhou; Zhonghua Liu

doi:10.3390/toxins10090358

Engineering Gain-of-Function Analogues of the Spider Venom Peptide HNTX-I, A Potent Blocker of the hNa_V1.7 Sodium Channel

Toxins (Basel). 2018 Sep 4;10(9):358. doi: 10.3390/toxins10090358.

Authors

Yunxiao Zhang¹, Qiuchu Yang², Qingfeng Zhang³, Dezheng Peng⁴, Minzhi Chen⁵, Songping Liang⁶, Xi Zhou⁷, Zhonghua Liu⁸

Affiliations

¹ The National & Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha 410081, Hunan, China. Zhangyx0954@163.com.
² The National & Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha 410081, Hunan, China. QQQQiuchu@126.com.
³ The National & Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha 410081, Hunan, China. feng122436ab@126.com.
⁴ The National & Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha 410081, Hunan, China. pdz920217@sina.com.
⁵ The National & Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha 410081, Hunan, China. chenmz@hunnu.edu.cn.
⁶ The National & Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha 410081, Hunan, China. liangsp@hunnu.edu.cn.
⁷ The National & Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha 410081, Hunan, China. xizh@hunnu.edu.cn.
⁸ The National & Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha 410081, Hunan, China. liuzh@hunnu.edu.cn.

Abstract

Pain is a medical condition that interferes with normal human life and work and reduces human well-being worldwide. The voltage-gated sodium channel (VGSC) human Na_V1.7 (hNa_V1.7) is a compelling target that plays a key role in human pain signaling. The 33-residue peptide µ-TRTX-Hhn2b (HNTX-I), a member of Na_V-targeting spider toxin (NaSpTx) family 1, has shown negligible activity on mammalian VGSCs, including the hNa_V1.7 channel. We engineered analogues of HNTX-I based on sequence conservation in NaSpTx family 1. Substitution of Asn for Ser at position 23 or Asp for His at position 26 conferred potent activity against hNa_V1.7. Moreover, multiple site mutations combined together afforded improvements in potency. Ultimately, we generated an analogue E1G⁻N23S⁻D26H⁻L32W with >300-fold improved potency compared with wild-type HNTX-1 on hNa_V1.7 (IC₅₀ 0.036 ± 0.007 µM). Structural simulation suggested that the charged surface and the hydrophobic surface of the modified peptide are responsible for binding affinity to the hNa_V1.7 channel, while variable residues may determine pharmacological specificity. Therefore, this study provides a profile for drug design targeting the hNa_V1.7 channel.

Keywords: HNTX-I; NaV1.7; engineering; spider venom; toxin; voltage-gated sodium channels.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Gain of Function Mutation
HEK293 Cells
Humans
Models, Molecular
NAV1.7 Voltage-Gated Sodium Channel / physiology*
Peptides* / chemistry
Peptides* / genetics
Peptides* / pharmacology
Sodium Channel Blockers* / chemistry
Sodium Channel Blockers* / pharmacology
Spider Venoms*

Substances

NAV1.7 Voltage-Gated Sodium Channel
Peptides
SCN9A protein, human
Sodium Channel Blockers
Spider Venoms