Reductive glutaraldehydation of amine groups for identification of protein N-termini
Reductive glutaraldehydation of amine groups for identification of protein N-termini
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DOI：
10.1021/pr800224v
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发表时间：
2008-09-01
影响因子：
4.4
通讯作者：
Deng, Haiteng
中科院分区：
文献类型：
--
作者：
Russo, Allison;Chandramouli, Nagarajan;Deng, Haiteng
在本工作中，蛋白质和肽与戊二醛的还原烷基化（还原戊二醛）的报告。该反应是高效的，并在N-末端以及赖氨酸残基的侧链处形成哌啶。在溶液中或在凝胶中，在不到15分钟的还原戊二醛化实现了蛋白胺的完全修饰。戊二醛修饰的肽显示增强的强度在质谱中，并显示更高的保留时间在反相色谱法相比，未修饰的肽。戊二醛修饰的蛋白质和肽的碎片化产生a1碎片离子，在MS/MS光谱中具有增强的强度。因此，一种基于还原性戊二醛和LC-MS/MS分析的方法已经被开发用于确定具有游离N-末端的蛋白质的N-末端残基。
In the present work, reductive alkylation of proteins and peptides with glutaraldehyde (reductive glutaraldehydation) is reported. The reaction is highly efficient and forms piperidine at the N-terminus as well as the side chain of lysine residues. The complete modification of protein amines was achieved by reductive glutaraldehydation in solution or in the gel in less than 15 min. The glutaraldehyde-modified peptides display an enhanced intensity in mass spectra and show higher retention time in reversed phase chromatography in comparison to unmodified peptides. Fragmentation of glutaraldehyde-modified proteins and peptides generates a1 fragment ions with enhanced intensity in MS/MS spectra. Thus, a method based on reductive glutaraldehydation and LC-MS/MS analysis has been developed to determine the N-terminal residue of proteins with free N-termini.