Foettinger A, Leitner A, Lindner W
Christian Doppler Laboratory for Molecular Recognition Materials, Institute of Analytical Chemistry, University of Vienna, Vienna, Austria.
J Chromatogr A. 2005 Jun 24;1079(1-2):187-96. doi: 10.1016/j.chroma.2005.03.038.
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.
本文介绍了一种通过固相捕获和释放技术从混合物中选择含精氨酸肽的方法。该方法基于在碱性条件下，精氨酸的胍基与2,3-丁二酮和苯硼酸进行共价修饰。使用固定化苯硼酸的聚合材料，含精氨酸的肽可以捕获在固体支持物上，而不含精氨酸的肽不会共价结合，可以被洗去。最后，由于反应的可逆性，含精氨酸的肽可以再次从硼酸珠上裂解下来。然后通过液相色谱-串联质谱对回收的肽进行分析。针对不含精氨酸肽的非特异性结合以及选择步骤后标签的定量裂解，使用模型肽对该方法进行了优化。使用适当的方案，通过三种标准蛋白质混合物的胰蛋白酶消化物成功测试了该方法对更复杂样品的适用性。
