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目的:利用诱导碰撞解离串联质谱技术,获得富含半胱氨酸毒素多肽α-芋螺毒素(α-CTx)LtIA的裂解规律,为分析未知天然α-芋螺毒素肽的氨基酸序列结构提供参比和方法借鉴。方法:通过Fmoc固相合成,色谱分离获得高纯度α-CTx LtIA还原型线性肽,然后利用离子阱-飞行时间质谱进行MS和MS/MS分析,同时优化碰撞条件,分析产生的碎片离子峰特点。结果:利用Fmoc固相合成法成功合成了α-CTx LtIA线性肽,通过串联质谱分析母离子峰,当碰撞能量和气体设置为50%时,气体毒素肽b离子碎片离子较多,且丰度最高。结论:利用IT-TOF-MSMS,设置好串联质谱分析条件,可以最大限度获得多肽序列结构信息,为新的芋螺毒素序列分析提供支持。
OBJECTIVE: To study the cleavage pattern of α-CTX LtIA, a cysteine toxin-rich peptide, by means of collision-dissociation tandem mass spectrometry and to provide an insight into the amino acid sequence structure of an unknown natural α-conotoxin peptide References and methods of reference. Methods: The high purity α-CTx LtIA reduced linear peptide was obtained by Fmoc solid-phase synthesis and chromatographic separation. The MS and MS / MS analysis were performed by ion trap-time of flight mass spectrometry. The collision conditions were optimized and the fragment ion peaks were analyzed . Results: The α-CTx LtIA linear peptide was successfully synthesized by Fmoc solid phase synthesis. The parent ion peak was analyzed by tandem mass spectrometry. When the collision energy and gas concentration were set to 50%, the gas ion peptide b ion fragment ions were more abundant, and the abundance highest. Conclusion: By using IT-TOF-MSMS, tandem mass spectrometry conditions can be set to obtain the maximum sequence information of polypeptide sequence and provide support for the new conotoxin sequence analysis.