Triazolecarbaldehyde Reagents for One‐Step N‐Terminal Protein Modification

Abstract

Site-specific modification of peptides and proteins is ak ey aspect of protein engineering.W ed eveloped am ethod for modification of the Nterminus of proteins using 1H-1,2,3-triazole-4-carbaldehyde( TA4C) derivatives, which can be prepared in one step. The N-terminal specific labelingo fb ioactive peptides and proteins with the TA4C derivatives proceeds under mild reactionc onditions in excellent conversion (angiotensinI: 92 %, ribonuclease A: 90 %). This method enables site-specific conjugation of various functional molecules such as fluorophores,b iotin, and polyethylene glycol attached to the triazole ring to the Nterminus. Furthermore, af unctional molecule modified with ap rimary amine moiety can be directly converted into aT A4C derivativethrough aDimroth rearrangement reaction with 1-(4-nitrophenyl)-1H-1,2,3-triazole-4-carbaldehyde. This methodc an be used to obtain N-terminal-modifiedp roteins via only two steps:1 )convenient preparation of aT A4C derivativew ith af unctional group and 2) modification of the Nterminus of the protein with the TA4C derivative.

Profiling the Proteome-Wide Selectivity of Diverse Electrophiles

“…Although several lysine-directed probes like STP-, ArSq-and EBA-alkyne also allow monitoring of N-termini, selective chemistry is highly desirable. Here, carboxaldehydes of electron-poor heteroaromatics have been previously applied to modify proteins (TCA- 49 and PCA-alkyne, 50 Fig. 4f).…”

Herding cats: Label-based approaches in protein translocation through nanopore sensors for single-molecule protein sequence analysis

“…More recently, other groups have also performed similar or revised approaches for N-terminal specific modification ( Chen et al., 2017 ; Deng et al., 2020 ; Inoue et al., 2019 ; Li et al., 2018 ; Onoda et al., 2020 ). In 2019, Inoue et al.…”

“…In 2020, Onoda et al. developed a more general strategy for one-step modification of the protein N-terminus using triazolecarbaldehyde reagents ( Onoda et al., 2020 ). In principle, this method is applicable for protein modification with any functional molecule containing a primary amine.…”

1-(4-Nitrophenyl)-1H-1,2,3-Triazole-4-carbaldehyde: Scalable Synthesis and Its Use in the Preparation of 1-Alkyl-4-Formyl-1,2,3-triazoles

“…4-Formyl-1,2,3-triazoles, also known as 1,2,3-triazole-4-carbaldehydes, are widely present in the literature, mainly due to the synthetic versatility of the formyl group [1][2][3][4][5][6][7]. In the field of medicinal chemistry [7], triazole-4-carbaldehydes have been prepared as intermediates in the synthesis of anticancer [8][9][10], antifungal [11,12], antituberculosis [13][14][15], anti-inflammatory [10,16], antidiabetic [17], and even bioimaging agents [18].…”

“…As a consequence, FNPT 1a was put forward as a preferred reagent for the synthesis of 1-subsituted 4-formyltriazoles and can be seen as a safe alternative for diazomalonaldehyde [14,35]. Onoda et al applied this strategy for the preparation of 1-alkyl-4-formyltriazoles 7 that could be used for the N-terminal modification of proteins [1]. Interestingly, a procedure was developed starting from FNPT 1a and primary amines that allowed for protein modification in excellent conversions without isolating the targeted aldehydes 7.…”