Liu Jiongyu, Jiang Jianping
Chengdu Institute of Biology, Chinese Academy of Sciences, PO Box 416, Chengdu, 610041, China.
Methods Mol Biol. 2017;1548:89-99. doi: 10.1007/978-1-4939-6737-7_7.
In the past decades, a great amount of antimicrobial peptides (AMPs) has been discovered, the structure identification of which relies heavily on de novo sequencing by Edman degradation or mass spectrometry. Here we outline the basic procedures for the exact mass measurement approaches that use off-line low-energy CID ESI Qq-TOF MS/MS in positive-ion mode, which is typically applied to de novo sequencing of peptides, to elucidate the structure of AMPs. Ambiguity I/L and partial sequence order were elucidated by Edman degradation or/and structural similarity analysis to known sequence. The approaches can determine the structure of peptides composed of as much as 38 amino acids in our practice.
在过去几十年中，人们发现了大量抗菌肽（AMPs），其结构鉴定在很大程度上依赖于通过埃德曼降解或质谱进行的从头测序。在此，我们概述了使用离线低能量碰撞诱导解离电喷雾串联四极杆飞行时间质谱（ESI Qq-TOF MS/MS）在正离子模式下进行精确质量测量的基本程序，该方法通常用于肽的从头测序，以阐明抗菌肽的结构。通过埃德曼降解或/和与已知序列的结构相似性分析来阐明模糊的I/L和部分序列顺序。在我们的实践中，这些方法可以确定由多达38个氨基酸组成的肽的结构。
