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Copper‐Assisted Click Reactions for Activity‐Based Proteomics: Fine‐Tuned Ligands and Refined Conditions Extend the Scope of Application
Author(s) -
Rudolf Georg C.,
Sieber Stephan A.
Publication year - 2013
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201300551
Subject(s) - click chemistry , bioconjugation , combinatorial chemistry , chemistry , azide , cycloaddition , tris , 1,3 dipolar cycloaddition , cationic polymerization , peptide , organic chemistry , biochemistry , catalysis
Abstract Copper‐catalysed alkyne–azide 1,3‐dipolar cycloaddition (CuAAC) is the predominantly used bioconjugation method in the field of activity‐based protein profiling (ABPP). Several limitations, however, including conversion efficiency, protein denaturation and buffer compatibility, restrict the scope of established procedures. We introduce an ABPP customised click methodology based on refined CuAAC conditions together with new accelerating copper ligands. A screen of several triazole compounds revealed the cationic quaternary {3‐[4‐({bis[(1‐ tert ‐butyl‐1 H ‐1,2,3‐triazol‐4‐yl)methyl]amino}methyl)‐1 H ‐1,2,3‐triazol‐1‐yl]propyl}trimethylammonium trifluoroacetate (TABTA) to be a superior ligand. TABTA exhibited excellent in vitro conjugation kinetics and optimal ABPP labelling activity while almost exclusively preserving the native protein fold. The application of this CuAAC‐promoting system is amenable to existing protocols with minimal perturbations and is even compatible with previously unusable buffer systems such as Tris ⋅ HCl.