One‐Step Azolation Strategy for Site‐ and Chemo‐Selective Labeling of Proteins with Mass‐Sensitive Probes

Abstract

The chemical modification of proteins in as iteselective manner leads to many advances in various scientific fields.T he major challenges with conventional N-terminal bioconjugation techniques are the lacko fu niversal sequence compatibility and poor mass-detection sensitivity of the resulting bioconjugates.T his approach efficiently analyzes proteolytic fragments and native proteins in ac omplex mixture. Multiple chemical steps are usually required for the siteselective synthesis of bioconjugates with enhanced massdetection sensitivity.W epresent asingle-step,versatile strategy for the selective modification of protein N-termini with mass boosters.T he chemical tag enhances the peptide detection by multiple orders thus leading to the unambiguous analysis of the resulting bioconjugates.W ed emonstrate that tagging proteolytic fragments with mass sensitivity probes in ac omplex mixture improves the detection of resulting bioconjugates.

An overview of chemo- and site-selectivity aspects in the chemical conjugation of proteins

“…2-Methylthio oxazolines 196 have also been applied to the site-selective conjugation of native proteins with almost complete conversion in several cases [238]. In addition, the oxazoline group helped to increase ionization of peptide fragments by mass spectrometry and enhance the sensitivity of the method, making this conjugation strategy valuable for proteomics studies even though it was only limited to relatively low molecular-weight proteins (i.e.…”