Solid-phase capture and release of arginine peptides by selective tagging and boronate affinity chromatography
Solid-phase capture and release of arginine peptides by selective tagging and boronate affinity chromatography
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DOI：
10.1016/j.chroma.2005.03.038
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发表时间：
2005-06-24
影响因子：
4.1
通讯作者：
Lindner, W
中科院分区：
文献类型：
--
作者：
Foettinger, A;Leitner, A;Lindner, W
提出了一种用固相捕获与释放技术从混合物中选择含精氨酸肽的方法。该方法是在碱性条件下用2,3-丁二酮和苯硼酸对精氨酸的胍基进行共价修饰。使用固定苯硼酸的聚合物材料，精氨酸肽可以被捕获在固体载体上，而无精氨酸肽不是共价结合的，可以被冲走。最后，由于反应的可逆性，精氨酸肽可以再次从硼酸珠中分离出来。然后用液相色谱-串联质谱法分析回收的肽。利用模型肽对该方法进行了优化，考虑了无精氨酸肽的非特异性结合和选择步骤后标记的定量切割。使用适当的协议，对更复杂的样品的适用性成功地测试了三种标准蛋白质的混合物的胰蛋白酶消化。(c) 2005 Elsevier B.V.版权所有
A method for the selection of arginine-containing peptides from a mixture by a solid phase capture and release technique is presented. The method is based on the covalent modification of the guanidine group of arginine with 2,3-butanedione and phenylboronic acid under alkaline conditions. Using polymeric materials with immobilised phenylboronic acid the arginine-peptides can be captured on a solid support while arginine-free peptides are not covalently bound and can be washed away. Finally, the arginine-peptides can be cleaved again from the boronic acid beads due to the reversibility of the reaction. The recovered peptides are then analysed by liquid chromatography-tandem mass spectrometry. The method was optimised with model peptides with regard to the non-specific binding of arginine-free peptides and quantitative cleavage of the label after the selection step. Using an adequate protocol, the applicability towards more complex samples was successfully tested with a tryptic digest of a mixture of three standard proteins. (c) 2005 Elsevier B.V. All rights reserved.