An in situ combinatorial methodology to synthesize and screen chemical probes

Abstract

Linking of reactive groups and ligands by imine chemistry provides chemical probes that label proteins of interest.

Development of an 18F-Labeled Irreversible Inhibitor of Transglutaminase 2 as Radiometric Tool for Quantitative Expression Profiling in Cells and Tissues

“…5 Nevertheless, also enzymes that do not rely on such intermediates and even proteins without catalytic activity can potentially be monitored in this way by targeting noncatalytic nucleophilic residues in these proteins. 6,7 However, the most straightforward approach for the design of activity-based probes is based on irreversible enzyme inhibitors containing electrophilic warheads that are capable of stable and selective covalent bonding to nucleophilic residues in the enzyme's active site. 8,9 The reporter tag, however, should preferably not compromise the inhibitory activity, the selectivity profile, and the membrane permeability of the original inhibitor molecule.…”

Profiling Sulfur(VI) Fluorides as Reactive Functionalities for Chemical Biology Tools and Expansion of the Ligandable Proteome

“…The reversible affinity of the fragments were typically beyond the limit of the assay (KI < 5 µM), consistent with previous reports of aryl primary sulfonamides showing submicromolar affinity for CAII (Figure 4E, 4F). 41,42 The rates of covalent modification (kinact) showed little correlation with the intrinsic reactivity of the S VI -F electrophile (Figure 4G). The results showed some correlation for a given fragment series, e.g.…”

Target identification of anticancer natural products using a chemical proteomics approach

“… 30 Other strategies like in situ combinatorial probe synthesis, label based approach also applicable to improve A f BP. 31 Sometimes it is spectacular that A f BP enabling drugs susceptible to recognize protein in complex proteome as low as in ppm label. 31 However, when the drug is designed as an A f BP, it must be functionalized to immobilize by covalently attached agarose or magnetic bead during the anchoring process, pharmacological activity of drug should not be disturbed.…”

“… 31 Sometimes it is spectacular that A f BP enabling drugs susceptible to recognize protein in complex proteome as low as in ppm label. 31 However, when the drug is designed as an A f BP, it must be functionalized to immobilize by covalently attached agarose or magnetic bead during the anchoring process, pharmacological activity of drug should not be disturbed. Subsequently, this immobilized probe complex was incubated with extracted proteins ( Fig.…”