Mikami Toshiyuki, Takao Toshifumi
Laboratory of Protein Profiling and Functional Proteomics, Institute for Protein Research, Osaka University, Yamadaoka 3-2, Suita, Osaka 565-0871, Japan.
Anal Chem. 2007 Oct 15;79(20):7910-5. doi: 10.1021/ac071294a. Epub 2007 Sep 14.
We developed a method for selective isolation of N-blocked peptides from a complex mixture such as an enzymatic digest of a protein. The approach is based on a newly designed isocyanate-resin (resin-NCO), which specifically reacts with alpha-amino or imino groups. This method, then, permits the isolation of N-blocked peptides, even those containing Lys, from a peptide mixture as intact forms by trapping N-free peptides via covalent bonding to the resin-NCO. The present study demonstrates the performance of this method for the selective isolation of N-blocked peptides by applying it to several peptide mixtures, including proteolytic digests.
我们开发了一种从复杂混合物(如蛋白质的酶解产物)中选择性分离N-封端肽的方法。该方法基于一种新设计的异氰酸酯树脂(树脂-NCO),它能与α-氨基或亚氨基特异性反应。然后,这种方法可以通过与树脂-NCO共价结合捕获无N肽,以完整形式从肽混合物中分离出N-封端肽,甚至是那些含有赖氨酸的肽。本研究通过将该方法应用于包括蛋白水解消化物在内的几种肽混合物,展示了其用于选择性分离N-封端肽的性能。