期刊
JOURNAL OF CHROMATOGRAPHY A
卷 1079, 期 1-2, 页码 187-196出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2005.03.038
关键词
arginine-peptides; phenylboronic acid beads; affinity enrichment; tryptic digest; mass spectrometry
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.
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