试剂
化学
氰醇钠
小学(天文学)
甲基化
赖氨酸
还原胺化
质谱法
组合化学
甲醛
同位素标记
氘
色谱法
有机化学
生物化学
氨基酸
DNA
物理
量子力学
天文
催化作用
作者
Alexandre K. Tashima,Leandro M. Castro,Lloyd D. Fricker
出处
期刊:Methods in molecular biology
日期:2024-01-01
卷期号:: 109-124
标识
DOI:10.1007/978-1-0716-3646-6_6
摘要
A number of different approaches have been used for quantitative peptidomics. In this protocol, we describe the method in which peptides are reacted with formaldehyde and sodium cyanoborohydride, which converts primary and secondary amines into tertiary amines. By using different combinations of regular reagents, deuterated reagents (2H), and reagents containing deuterium and 13C, it is possible to produce five isotopically distinct forms of the methylated peptides, which can be quantified by mass spectrometry. Peptides with free N-termini that are primary amines incorporate two methyl groups using this procedure, which differ by 2 Da for each of the five isotopic combinations. Peptides that contain unmodified lysine residues incorporate additional pairs of methyl groups, leading to larger mass differences between isotopic forms. The reagents are commercially available, relatively inexpensive, and chemically stable.
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