α-芋螺毒素TxIB氧化折叠路径研究

Study on oxidative folding pathway ofα-conotoxin TxIB

目的阐明TxIB线性肽通过一部氧化法,在体外被氧化折叠形成含有两对二硫键活性肽的具体路径,为后续大量人工合成该活性肽的方法研究提供基础。方法采用N-9芴甲氧羰基(Fmoc)固相合成法人工合成TxIB线性肽,用高效液相和质谱分析法,监测研究TxIB一步氧化折叠过程中所经历的路径。结果 TxIB氧化路径经历了1对二硫键(1SS)、2对二硫键(X-2SS)折叠中间体后,最终形成稳定的含有2对二硫键(N-2SS)的氧化型TxIB。结论芋螺毒素TxIB折叠路径与前期研究的水蛭素氧化折叠路径一致。

Objective To elucidate one-step oxidative folding pathway of linear TxIB peptide,and to give aguidance to synthesize peptide TxIB in large amounts.Methods Linear TxIB was synthesized by solidphase synthesis method,and the pathway of oxidative folding was monitored and determined by reverse phase high performance liquid chromatography and mass spectrometry analysis.Results The folding intermediates of reduced TxIB had one disulfide bond（1SS）,two disulfide bonds（X-2SS）,and formed the active TxIB peptide with two stable disulfide bonds（N-2SS）finally.Conclusion The pathway of oxidative folding of peptide TxIB exhibited a high degree of similarity to hirudin.