CAJ | 학술논문

目的：合成难溶的新4/3型α-芋螺毒素Eb1.3突变体Eb1.3[ΔR1，W13M]，并初步测定其与烟碱型乙酰胆碱受体亚型的结合作用。方法：采用Fmoc-固相法合成线性肽Eb1.3[ΔR1，W13M]，通过空气氧化折叠和磺化产物折叠获得含二硫键的折叠产物，利用两步折叠法测定其二硫键连接方式，双电极电压钳技术检测其药理活性。结果：Eb1.3[ΔR1，W13M]线性肽经折叠生成的主产物Ⅰ的二硫键排列方式为少见的C1-C4、C2-C3，而非典型的C1-C3、C2-C4，线性肽磺化后再经GSH交换可加速折叠，10μmol/L的产物Ⅰ对乙酰胆碱受体α3β2亚型的抑制率为28.97%±8.44%。结论：新4/3型α-芋螺毒素Eb1.3突变体Eb1.3[ΔR1，W13M]形成非典型的二硫键C1-C4、C2-C3，产物Ⅰ对乙酰胆碱受体α3β2亚型具有一定的结合活性。
목적：합성난용적신4/3형α-우라독소Eb1.3돌변체Eb1.3[ΔR1，W13M]，병초보측정기여연감형을선담감수체아형적결합작용。방법：채용Fmoc-고상법합성선성태Eb1.3[ΔR1，W13M]，통과공기양화절첩화광화산물절첩획득함이류건적절첩산물，이용량보절첩법측정기이류건련접방식，쌍전겁전압겸기술검측기약리활성。결과：Eb1.3[ΔR1，W13M]선성태경절첩생성적주산물Ⅰ적이류건배렬방식위소견적C1-C4、C2-C3，이비전형적C1-C3、C2-C4，선성태광화후재경GSH교환가가속절첩，10μmol/L적산물Ⅰ대을선담감수체α3β2아형적억제솔위28.97%±8.44%。결론：신4/3형α-우라독소Eb1.3돌변체Eb1.3[ΔR1，W13M]형성비전형적이류건C1-C4、C2-C3，산물Ⅰ대을선담감수체α3β2아형구유일정적결합활성。
Objective: To synthesize the new insoluble 4/3 type α-conotoxin Eb1.3 mutant EB1.3[ΔR1,W13M] and determinate its binding ability to neuronal nicotinic acetylcholine receptor(nAChR) subtypes. Methods: The Eb1.3[ΔR1,W13M] linear peptide was synthesized by Fmoc-solid phase synthesis. The air oxidative folding and sulfonated modification folding were used to get aim products. The two-step oxidative folding method was then em?ployed to determine the disulfide bridge connections. Two electrode voltage clamp technique was used to deter?mine its binding to neuronal nicotinic acetylcholine receptor subtypes. Results: The disulfide-bridge framework of the main folding product of Eb1.3[ΔR1,W13M] linear peptide is a rare C1-C4,C2-C3 but not the typical C1-C3, C2-C4. The folding of sulfonated liner peptide of Eb1.3[ΔR1,W13M] was fast in the presence of reduced and oxi?dized glutathione. The inhibition ratio of Eb1.3[ΔR1,W13M] with nAChR α3β2 subtype is 28.97%±8.44% at 10μmol/L. Conclusion: Eb1.3[ΔR1,W13M] has a unique disulfide-bridge framework C1-C4,C2-C3 and is active to nAChR α3β2 subtype.