| 范瑜轩,何全阔,吴勇,等.Sortase A环化α-芋螺毒素LvIA研究[J].中国海洋药物,2026,45(1):1-8. |
| Sortase A环化α-芋螺毒素LvIA研究 |
| Study on Sortase A cyclization of α-conotoxin LvIA |
| 投稿时间：2024-04-30 修订日期：2024-06-15 |
| DOI：10.13400/j.cnki.cjmd.2026.01.012 |
| 中文关键词: Sortase A α-芋螺毒素LvIA 环化改造 稳定性 α3β2 nAChR |
| English Keywords:Sortase A α-conotoxin LvIA cyclization modification stability α3β2 nAChR |
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| 中文摘要: |
| 摘要：目的 α-芋螺毒素LvIA能够选择性抑制α3β2型乙酰胆碱受体（nAChR），可作为研究α3β2的分子探针或作为研究新型镇痛药物的候选分子。本研究的目的是利用Sortase A（Srt A）环化LvIA,并评估cLvIA在不同环境下的稳定性。方法 本研究利用基因工程方法表达环化酶Srt A，利用环化酶和Fmoc固相合成法合成首尾环化的环肽cLvIA。在此基础上，利用反相-超高压液相色谱（RP-UPLC）方法评估cLvIA稳定性，蛙卵膜片钳方法检测其抑制α3β2 nAChR活性。结果 利用Srt A可以成功地对LvIA进行环化修饰，cLvIA在血清、蛋白酶K和高温条件下表现出较高的稳定性。电生理活性初步评估表明，在10 μmol·L-1浓度下，cLvIA能够阻断90%以上的受体活性，初步表明cLvIA保持了原有的活性。本研究表明利用Srt A可以作为芋螺毒素LvIA环化修饰有效手段，为其提高稳定性提供了重要的修饰策略，也为其他芋螺毒素环化研究提供重要指导。 |
| English Summary: |
| ABSTRACT:Objective Conotoxin α-LvIA exhibits high selectivity in inhibiting the α3β2 subtype of nicotinic acetylcholine receptor (nAChR), making it a potential candidate for novel analgesic drugs. This study aims to cyclize LvIA using Sortase A (Srt A) and evaluate the stability of cLvIA in different environments. Methods Sortase A was expressed using genetic engineering methods to cyclize LvIA, and cyclic peptide cLvIA was synthesized via solid-phase synthesis using Fmoc chemistry. Subsequently, the stability of cLvIA was assessed using reverse-phase ultra-high performance liquid chromatography (RP-UPLC), and its inhibition of α3β2 nAChR activity was evaluated using frog oocyte electrophysiology. Results Successful cyclization of LvIA was achieved using Srt A, and cLvIA exhibited high stability under serum, proteinase K, and high-temperature conditions. Preliminary electrophysiological activity evaluation indicates that cLvIA, at a concentration of 10 μmol·L-1, can block over 90% of receptor activity, suggesting that cyclization maintains the original activity of LvIA. This study highlights the efficacy of Srt A-mediated cyclization in modifying LvIA, providing important insights for enhancing its stability and guiding further research on cyclization of other conotoxins. |
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