Solid-phase capture and release of arginine peptides by selective tagging and boronate affinity chromatography.
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.
Foettinger
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A
Christian Doppler Laboratory for Molecular Recognition Materials, Institute of Analytical Chemistry, University of Vienna, Vienna, Austria.
Leitner
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A
Lindner
W
W
D001120:Arginine / Q000737:chemistry*; D001897:Boronic Acids / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000379:methods*; D010455:Peptides / Q000302:isolation & purification*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000378:metabolism
D001897:Boronic Acids; D010455:Peptides; D001120:Arginine; D014357:Trypsin; C010686:benzeneboronic acid