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Multiplexed azido‐group isotopic capture (MAGIC) beads: Selective analysis of azido compounds using a propargyl‐based cleavable linker, a proof of concept
Author(s) -
McLellan Michelle,
Doyle Michael G. J.,
Bodnar Edward D.,
Lopez Paul G.,
Domalaon Ronald,
Roy Rini,
Cordova Katherine,
Schweizer Frank,
Perreault Hélène
Publication year - 2016
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.7744
Subject(s) - chemistry , azide , linker , click chemistry , peptide , combinatorial chemistry , propargyl , alkyne , mass spectrometry , peptide synthesis , matrix assisted laser desorption/ionization , cycloaddition , organic chemistry , chromatography , desorption , adsorption , catalysis , biochemistry , computer science , operating system
Rationale A cleavable linker is designed and synthesized for the selective capture of azide‐containing compounds. This article presents a proof of concept methodology involving the use of peptide‐functionalized aminopropyl silica, on which the peptide is constructed by solid‐phase peptide synthesis. Methods The peptide linker has L‐propargylglycine (Pra) at one terminal end to allow the conjugation of azide‐containing molecules by copper assisted azide alkyne cycloaddition, also known as click reaction. L‐Arginine (Arg) is placed just before Pra to permit the release of the captured product by tryptic cleavage. Three glycine (Gly) residues, as part of the linker, are appended to the silica bead to present a spacer section that allows efficient tryptic cleavage devoid of steric hindrance imposed by the bulky bead. The bead composition is Si‐O‐propyl‐NH‐Gly‐Gly‐Gly‐Arg‐Pra. Results This solid‐phase material can be used to capture and release azide‐functionalized compounds. The beads are first tested on three azido compounds, 2‐azido‐2‐deoxyglucose (ADG), BOC‐ p ‐azido‐Phe‐OH (BAzPhe), where BOC = tert ‐butoxycarbonyl, and tetraacetylated‐ N ‐azidomannosamine (Ac 4 ManNAz). Copper‐mediated click reaction conditions are used and released products are characterized by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF‐MS) and tandem MS (MS/MS). Conclusions This method allows easy identification of captured compounds based on mass and fragmentation analysis. Moreover, it is useful for the analysis of small azide‐containing compounds by MALDI‐TOF‐MS which may not be possible otherwise due to matrix interferences. The insertion of isotopically labeled Arg residues provides the possibility of multiplex analysis, from which the beads have been called MAGIC (for Multiplexed Azido‐Group Isotopic Capture). Copyright © 2016 John Wiley & Sons, Ltd.