Hoyt, E. A., Cal, P. M. S. D., Oliveira, B. L. & Bernardes, G. J. L. Contemporary approaches to site-selective protein modification. Nat. Rev. Chem. 3, 147–171 (2019).

Article  CAS  Google Scholar 

Dumontet, C., Reichert, J. M., Senter, P. D., Lambert, J. M. & Beck, A. Antibody–drug conjugates come of age in oncology. Nat. Rev. Drug Discov. 22, 641–661 (2023).

Article  CAS  PubMed  Google Scholar 

Ebrahimi, S. B. & Samanta, D. Engineering protein-based therapeutics through structural and chemical design. Nat. Commun. 14, 2411 (2023).

Article  CAS  PubMed  PubMed Central  Google Scholar 

Muttenthaler, M., King, G. F., Adams, D. J. & Alewood, P. E. Trends in peptide drug discovery. Nat. Rev. Drug Discov. 20, 309–325 (2021).

Article  CAS  PubMed  Google Scholar 

Rehm, F. B. H. et al. Asparaginyl ligases: new enzymes for the protein engineer’s toolbox. ChemBioChem 22, 2079–2086 (2021).

Article  CAS  PubMed  Google Scholar 

Harris, K. S. et al. Efficient backbone cyclization of linear peptides by a recombinant asparaginyl endopeptidase. Nat. Commun. 6, 10199 (2015).

Article  CAS  PubMed  PubMed Central  Google Scholar 

Yap, K. et al. An environmentally sustainable biomimetic production of cyclic disulfide-rich peptides. Green Chem. 22, 5002–5016 (2020).

Article  CAS  Google Scholar 

Yap, K. et al. Yeast-based bioproduction of disulfide-rich peptides and their cyclization via asparaginyl endopeptidases. Nat. Protoc. 16, 1740–1760 (2021).

Article  CAS  PubMed  Google Scholar 

Yang, R. et al. Engineering a catalytically efficient recombinant protein ligase. J. Am. Chem. Soc. 139, 5351–5358 (2017).

Article  CAS  PubMed  Google Scholar 

Rehm, F. B. H. et al. Site-specific sequential protein labeling catalyzed by a single recombinant ligase. J. Am. Chem. Soc. 141, 17388–17393 (2019).

Article  CAS  PubMed  PubMed Central  Google Scholar 

Zhou, Y. et al. A chemoenzymatic approach to produce a cyclic analogue of the analgesic drug MVIIA (ziconotide). Angew. Chem. Int. Ed. 62, e202302812 (2023).

Article  CAS  Google Scholar 

Le, T., Zhang, D., Martini, R. M., Biswas, S. & van der Donk, W. A. Use of a head-to-tail peptide cyclase to prepare hybrid RiPPs. Chem. Commun. 60, 6508–6511 (2024).

Article  CAS  Google Scholar 

Du, J. et al. A bifunctional asparaginyl endopeptidase efficiently catalyzes both cleavage and cyclization of cyclic trypsin inhibitors. Nat. Commun. 11, 1575 (2020).

Article  CAS  PubMed  PubMed Central  Google Scholar 

Li, C. Y. et al. Cystine knot peptides with tuneable activity and mechanism. Angew. Chem. Int. Ed. 61, e202200951 (2022).

Article  CAS  Google Scholar 

Hemu, X. et al. Structural determinants for peptide-bond formation by asparaginyl ligases. Proc. Natl Acad. Sci. USA 116, 11737–11746 (2019).

Article  CAS  PubMed  PubMed Central  Google Scholar 

Nguyen, G. K. T. et al. Butelase 1 is an Asx-specific ligase enabling peptide macrocyclization and synthesis. Nat. Chem. Biol. 10, 732–738 (2014).

Article  CAS  PubMed  Google Scholar 

Nguyen, G. K. T. et al. Butelase-mediated cyclization and ligation of peptides and proteins. Nat. Protoc. 11, 1977–1988 (2016).

Article  CAS  PubMed  Google Scholar 

Tang, T. M. S. et al. Use of an asparaginyl endopeptidase for chemoenzymatic peptide and protein labeling. Chem. Sci. 11, 5881–5888 (2020).

Article  CAS  PubMed  PubMed Central  Google Scholar 

Rehm, F. B. H., Tyler, T. J., Yap, K., Durek, T. & Craik, D. J. Improved asparaginyl-ligase-catalyzed transpeptidation via selective nucleophile quenching. Angew. Chem. Int. Ed. 60, 4004–4008 (2021).

Article  CAS  Google Scholar 

Chen, J. et al. Improvement of transpeptidative protein ligation via selective nucleophile quenching using 2-cyano-6-hydroxybenzothiazole. Org. Lett. 27, 3476–3481 (2025).

Article  CAS  PubMed  Google Scholar 

Xia, Y. et al. A cascade enzymatic reaction scheme for irreversible transpeptidative protein ligation. J. Am. Chem. Soc. 145, 6838–6844 (2023).

Article  CAS  PubMed  Google Scholar 

Nguyen, G. K. T., Cao, Y., Wang, W., Liu, C. F. & Tam, J. P. Site-specific N-terminal labeling of peptides and proteins using butelase 1 and thiodepsipeptide. Angew. Chem. Int. Ed. 54, 15694–15698 (2015).

Article  CAS  Google Scholar 

Rehm, F. B. H., Tyler, T. J., de Veer, S. J., Craik, D. J. & Durek, T. Enzymatic C-to-C protein ligation. Angew. Chem. Int. Ed. 61, e202116672 (2022).

Article  CAS  Google Scholar 

de Veer, S. J., Zhou, Y., Durek, T., Craik, D. J. & Rehm, F. B. H. Tertiary amide bond formation by an engineered asparaginyl ligase. Chem. Sci. 15, 5248–5255 (2024).

Article  CAS  PubMed  PubMed Central  Google Scholar 

Rehm, F. B. H. et al. Enzymatic C-terminal protein engineering with amines. J. Am. Chem. Soc. 143, 19498–19504 (2021).

Article  CAS  PubMed  Google Scholar 

Rehm, F. B. H. et al. Repurposing a plant peptide cyclase for targeted lysine acylation. Nat. Chem. 16, 1481–1489 (2024).

Article  CAS  PubMed  PubMed Central  Google Scholar 

de Veer, S. J., Craik, D. J. & Rehm, F. B. H. Highly efficient transpeptidase-catalyzed isopeptide ligation. J. Am. Chem. Soc. 147, 557–565 (2025).

Article  CAS  PubMed  Google Scholar 

Zhou, Y. et al. Asparaginyl ligases with engineered substrate specificity for controlled, sequential transpeptidation reactions. J. Am. Chem. Soc. 147, 21824–21831 (2025).

Article  CAS  PubMed  PubMed Central  Google Scholar 

Hemu, X. et al. Turning an asparaginyl endopeptidase into a peptide ligase. ACS Catal 10, 8825–8834 (2020).

Article  CAS  Google Scholar 

Liu, C. C. & Schultz, P. G. Adding new chemistries to the genetic code. Annu. Rev. Biochem. 79, 413–444 (2010).

Article  CAS  PubMed  Google Scholar 

Boutureira, O. & Bernardes, G. J. Advances in chemical protein modification. Chem. Rev. 115, 2174–2195 (2015).

Article  CAS  PubMed  Google Scholar 

Glasgow, J. E., Salit, M. L. & Cochran, J. R. In vivo site-specific protein tagging with diverse amines using an engineered sortase variant. J. Am. Chem. Soc. 138, 7496–7499 (2016).

Article  CAS  PubMed  Google Scholar 

Weikart, N. D. & Mootz, H. D. Generation of site-specific and enzymatically stable conjugates of recombinant proteins with ubiquitin-like modifiers by the Cu(I)-catalyzed azide-alkyne cycloaddition. ChemBioChem 11, 774–777 (2010).

Article