Ultrafast and selective labeling of endogenous proteins using affinity-based benzotriazole chemistry

Abstract

Chemical modification of proteins is enormously useful for characterizing protein function in complex biological systems and for drug development. Selective labeling of native or endogenous proteins is challenging owing to...

Ligand‐Directed Chemistry for Protein Labeling for Affinity‐Based Protein Analysis

“…In‐depth mass‐spectrometric analyses confirmed that these modified amino acids are localized in the proximity of the active pocket of the POI but not inside, which resulted in POI labeling while retaining its functions. After the first success using tosylate, several cleavable electrophiles have been developed as reactive modules to date as shown in Figure 2A, including phenyl ester, [27,28] acyl imidazole, [29,30] dibromophenyl benzoate, [31] N ‐acyl‐ N ‐alkyl‐sulfonamide (these undergo acyl transfer), [32,33] N ‐sulfonyl pyridone (protein sulfonation), [34] sulfone azide, (Cu 2+ catalyzed diazotransfer on Lys) [35] N ‐sulfanylethylanilide (SEAlide) (environment responsive N‐to‐S acyl transfer followed by acyl transfer on the reactive amino acid), [36] O ‐nitrobenzoxadiazole ( O ‐NBD: SN aromatic reaction), [37–39] TCC probe (Nucleophilic addition to alkyne), [40] and others (Figure 2A and Figure S1) [41–44] . It is noted that each electrophile showed distinct amino acids selectivity in the labeling reaction and varied labeling kinetics.…”

“… Chemical electrophiles exploited for LD chemistry (A) [23–44] . Cyclization reaction‐based turn‐on probe (TCC probe) for covalent labeling of proteins (B) [40] .…”

“…After the first success using tosylate, several cleavable electrophiles have been developed as reactive modules to date as shown in Figure 2A, including phenyl ester, [27,28] acyl imidazole, [29,30] dibromophenyl benzoate, [31] N-acyl-N-alkyl-sulfonamide (these undergo acyl transfer), [32,33] N-sulfonyl pyridone (protein sulfonation), [34] sulfone azide, (Cu 2 + catalyzed diazotransfer on Lys) [35] Nsulfanylethylanilide (SEAlide) (environment responsive N-to-S acyl transfer followed by acyl transfer on the reactive amino acid), [36] O-nitrobenzoxadiazole (O-NBD: SN aromatic reaction), [37][38][39] TCC probe (Nucleophilic addition to alkyne), [40] and others (Figure 2A and Figure S1). [41][42][43][44] It is noted that each electrophile showed distinct amino acids selectivity in the labeling reaction and varied labeling kinetics. These progresses are valuable to expand the availability/flexibility of LDchem for targeting diverse POIs in a variety of research objectives.…”

“…As another recent example, Arttamangkul et al employed LDAI chemistry for visualizing endogenous μ-opioid receptors (GPCRs) in living neurons on rat and mouse brain slices without loss of receptor functions. [43] Stoddart et al also reported chemical labeling of adenosine A 2A receptor (another GPCRs) endogenously expressed on the surface of SK-BR-3 cells by LDchem. [39] These examples clearly exhibited the versatility and robustness of LDchems for non-invasive and selective modification of membrane-bound proteins under live cell conditions.…”

“…As another recent example, Arttamangkul et al . employed LDAI chemistry for visualizing endogenous μ‐opioid receptors (GPCRs) in living neurons on rat and mouse brain slices without loss of receptor functions [43] . Stoddart et al .…”