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Combination of continuous digestion by peptidase and spectral similarity comparisons for peptide sequencing
Author(s) -
Yang Chao,
Liu Jianhui,
Hu Yechen,
Dai Zhongpeng,
Liang Zhen,
Shan Yichu,
Zhang Lihua,
Zhang Yukui
Publication year - 2020
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.202000459
Subject(s) - peptide , peptide sequence , carboxypeptidase , chemistry , amino acid , opioid peptide , biochemistry , exopeptidase , residue (chemistry) , computational biology , biology , enzyme , gene , receptor , opioid
Peptide sequencing is critical to the quality control of peptide drugs and functional studies of active peptides. A combination of peptidase digestion and mass spectrometry technology is common for peptide sequencing. However, such methods often cannot obtain the complete sequence of a peptide due to insufficient amino acid sequence information. Here, we developed a method of generating full peptide ladders and comparing their MS 2 spectral similarities. The peptide ladders, of which each component was different from the next component with one residue, were generated by continuous digestion by peptidase (carboxypeptidase Y and aminopeptidase). Then, based on the characteristics of peptide ladders, complete sequencing was realized by comparing MS 2 spectral similarity of the generated peptide ladders. The complete amino acid sequences of bivalirudin, adrenocorticotropic hormone, and oxytocin were determined with high accuracy. This approach is beneficial to the quality control of drug peptides as well as the identification of novel bioactive peptides.