Triazolinedione protein modification: from an overlooked off-target effect to a tryptophan-based bioconjugation strategy

Decoene, Klaas W.; Unal, Kamil; Staes, An; Zwaenepoel, Olivier; Gettemans, Jan; Gevaert, Kris; Winne, Johan M.; Madder, Annemieke

doi:10.1039/d1sc06942j

Cited by 13 publications

(23 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Furthermore, the reaction was observed for otherwise inaccessible Trp residues, as this method is less sensitive for the surface exposition of the residues, compared to other methods. Another recent example was published in 2022 by Madder and coworkers who report that an overlooked side reaction with Trp during triazolinedione based tyrosine labeling, can be used for selective Trp modification at lower pH (pH 4) (Figure 7E) [129] . The reaction is fast (occurs within a few seconds), is performed at rt and showed 50–86 % conjugation for the recombinant proteins tested in this study.…”

Section: Hydrophobic Amino Acid Residuesmentioning

confidence: 56%

“…Another recent example was published in 2022 by Madder and coworkers who report that an overlooked side reaction with Trp during triazolinedione based tyrosine labeling, can be used for selective Trp modification at lower pH (pH 4) (Figure 7E). [129] The reaction is fast (occurs within a few seconds), is performed at rt and showed 50-86 % conjugation for the recombinant proteins tested in this study. It therefore constitutes a new strategy for selective Trp conjugation, however, it is only useful for proteins that can withstand reaction at pH 4.…”

Section: Tryptophanmentioning

confidence: 78%

“…[117,118,119,121,123,124,126,129,131,132] Tyr 3.30 (0.63) Not covered here. Review: [39, 40] Val 7.01 (1.18) -…”

mentioning

confidence: 99%

See 2 more Smart Citations

Chemical Conjugation to Less Targeted Proteinogenic Amino Acids

2022

View full text Add to dashboard Cite

Protein bioconjugates are in high demand for applications in biomedicine, diagnostics, chemical biology and bionanotechnology. Proteins are large and sensitive molecules containing multiple different functional groups and in particular nucleophilic groups. In bioconjugation reactions it can therefore be challenging to obtain a homogeneous product in high yield. Numerous strategies for protein conjugation have been developed, of which a vast majority target lysine, cysteine and to a lesser extend tyrosine. Likewise, several methods that involve recombinantly engineered protein tags have been reported. In recent years a number of methods have emerged for chemical bioconjugation to other amino acids and in this review, we present the progress in this area.

show abstract

Section: Hydrophobic Amino Acid Residuesmentioning

confidence: 56%

Section: Tryptophanmentioning

confidence: 78%

See 1 more Smart Citation

Chemical Conjugation to Less Targeted Proteinogenic Amino Acids

2022

View full text Add to dashboard Cite

show abstract

“…131 (D) Ene-like reaction with triazolinediones at acidic pH. 169 (E) Electrochemical bioconjugation using keto-ABNO and 4-oxo-TEMPO. 162 (F) Radical trifluoromethylation using trifluoromethanesulfinate (Langlois' reagent).…”

Section: Iodanesmentioning

confidence: 99%

Chemical modification of proteins – challenges and trends at the start of the 2020s

2023

View full text Add to dashboard Cite

show abstract

“…Accordingly, many TAD reagents have been prepared for protein bioconjugation studies, discriminating changes in protein structure, 46 and recent electrochemical oxidation methods have been developed to enable the in situ generation of N4-substituted TAD reagents from their corresponding N4-substituted urazole precursors. [47][48][49][50][51] The procedural ease of these developments encouraged us to examine scenarios where in situ generated N4-substituted TAD peptides could react with sequence-embedded Tyr residues to produce Tyr-linked cyclic peptides (Figure 1C). The method design was predicated on several objectives.…”

Section: Introductionmentioning

confidence: 99%

Chemoselective, oxidation-induced macrocyclization of tyrosine-containing peptides

Keyes

Mifflin

Austin

et al. 2023

Preprint

View full text Add to dashboard Cite

Inspired by Nature’s wide range of oxidation-induced modifications to install cross-links and cycles at tyrosine (Tyr) and other phenol-containing residue side chains, we report a Tyr-selective strategy for the preparation of Tyr-linked cyclic peptides. This approach leverages N4-substituted 1,2,4-triazoline-3,5-diones (TADs) as azo electrophiles that react chemoselectively with the phenolic side chain of Tyr residues to form stable C–N1-linked cyclic peptides. In the developed method, a precursor 1,2,4-triazolidine-3,5-dione moiety, also known as a urazole, is readily constructed at any free amine revealed on a solid-supported peptide. Once prepared, the N4-substituted urazole peptide is selectively oxidized using mild, peptide-compatible conditions to generate an electrophilic N4-substituted TAD peptide intermediate that reacts selectively under aqueous conditions with internal and terminal Tyr residues to furnish Tyr-linked cyclic peptides. The approach demonstrates good tolerance of native residue side chains and enables access to cyclic peptides ranging from 3- to 11-residues in size (16- to 38-atom-containing cycles). The identity of the installed Tyr-linkage, a stable covalent C–N1 bond, was characterized using NMR spectroscopy. Finally, we applied the developed method to prepare biologically active Tyr-linked cyclic pep-tides bearing the integrin-binding RGDf epitope.

show abstract

Triazolinedione protein modification: from an overlooked off-target effect to a tryptophan-based bioconjugation strategy

Abstract: Labelling of tyrosine residues in peptides and proteins has been reported to selectively occur via a 'tyrosine-click' reaction with triazolinedione reagents (TAD). However, we here demonstrate that TAD reagents are...

Cited by 13 publications

References 39 publications

Chemical Conjugation to Less Targeted Proteinogenic Amino Acids

Chemical Conjugation to Less Targeted Proteinogenic Amino Acids

Chemical modification of proteins – challenges and trends at the start of the 2020s

Chemoselective, oxidation-induced macrocyclization of tyrosine-containing peptides

Contact Info

Product

Resources

About