Development of a fluoride-responsive amide bond cleavage device that is potentially applicable to a traceable linker

Abstract

A fluoride-responsive (FR) amino acid that induces amide bond cleavage upon the addition of a fluoride was developed, and it was applied to a FR traceable linker. By the use of an alkyne-containing peptide as a model of an alkynylatd target protein of a bioactive compound, introduction of the FR traceable linker onto the peptide was achieved. Subsequent fluoride induced cleavage of the linker followed by labeling of the released peptide derivative was also conducted to examine the potential applicability of the FR traceable linker to the enrichment and labeling of alkynylated target molecules.

Amide Bond Activation of Biological Molecules

“…Otaka et al developed an auxiliary with special protecting groups (PGs), which is capable of forming a lactone with the carbonyl of an amide bond, resulting in the cleavage of the amide bond ( Table 4 ) [ 121 , 122 , 123 , 124 , 125 , 126 , 127 , 128 ]. Depending on the nature of the protecting groups, an amide bond cleavage can be initiated in peptides by using different responsive reagents for the deprotection of PGs ( Table 4 ).…”

“…Table 4 showed various PGs and the corresponding responsive reagents for their deprotection. The thiol responsive reagent was applied for the cleavage of the PNA/DNA complex using thiol-responsive protecting groups [ 121 , 122 , 123 , 124 , 125 , 126 , 127 , 128 ].…”

Development of Chemical Biology Tools Focusing on Peptide/Amide Bond Cleavage Reaction

“…3. Not only UV 13,14) butalsonear-IR(NIR)two-photonexcitation, 15) intracellular hypoxic environment, 16) thiol, 17,18) hydrogen peroxide, 19) fluoride anion, 20) and alkaline phosphatase 14) Akira Shigenaga can trigger peptide bond cleavage of Spr. Several Spr-containing peptides were already used to control function of peptides/ proteins in living cells.…”

Development of a Turn-On Fluorescent Traceable Linker Employing <i>N</i>-Sulfanylethylcoumarinyl Amide for the Enrichment and Visualization of Target Proteins

“…[17][18][19] Because the contamination of non-target proteins hampers the subsequent identification of target proteins, several research groups including ours were prompted to engage in the development of an advanced cleavable linker. [20][21][22][23][24][25][26] The new cleavable linkers were designed to selectively label a target protein for facile discrimination of the target from non-targets. These linkers enable label-based tracing of the target; therefore, we named this type of cleavable linker a "traceable linker."…”

“…Because phosphate activates the SEAlide unit from an amide to a thioester form, 28,29) the addition of a phosphate buffer containing reductant for the removal of a t-BuS group of the SEAlide moiety was necessary, and a labeling reagent with a cysteine moiety induced bioothogonal Native Chemical Ligation (NCL) 30) to elute a labeled target protein. In this protocol, an external labeling reagent is required, similar to the other traceable linkers, [21][22][23][24][25] and complete removal of the excess labeling reagent was sometimes complicated. When sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and subsequent fluorescence imaging of the eluent with the residual labeling reagent was examined, fluorescence caused by the labeling reagent was observed in the low molecular weight region.…”