Butelase-Mediated Ligation as an Efficient Bioconjugation Method for the Synthesis of Peptide Dendrimers

Abstract

Herein we report a novel enzymatic bioconjugation method to prepare peptide dendrimers. Under the catalysis of a newly discovered peptide ligase, butelase 1, peptide dendrimers of di-, tetra-, and octabranches were successfully synthesized using thiodepsipeptides as acyl donors for ligation with lysyl dendrimeric scaffolds. The efficient assembly of the highly clustered dendrimeric structure highlighted the versatility of butelase 1. We also showed that our synthetic antibacterial peptide dendrimers containing an RLYR motif are highly potent and broadly active against antibiotic-resistant strains.

Bioconjugation Approaches to Producing Subunit Vaccines Composed of Protein or Peptide Antigens and Covalently Attached Toll-Like Receptor Ligands

“…Further, it has been reported to be the fastest-acting ligase known to date, and has been used in various studies for the ligation or cyclization of peptides and proteins. 96,109,110 The required flanking sequences are also simple, e.g. an Asp/Asn-His-Val motif at the C-terminus of peptide 1 and a Xaa-Ile/Leu/Val/Cys motif at the N-terminus of peptide 2 (Figure 7; where Xaa is any amino acid except proline).…”

Natural Occurring and Engineered Enzymes for Peptide Ligation and Cyclization

“…Besides cyclization, Butelase-1 can be used for the modification of live cell bacterial surfaces (Bi et al, 2017), for the semi-synthesis of ubiquitin (Nguyen et al, 2015a), and to prepare large circular bacteriocins, the largest antimicrobial peptides known up to date (Hemu et al, 2016). Other possibilities are the preparation of peptide dendrimers using lysine derived scaffolds (Cao et al, 2016) or even the modification of proteins (Nguyen et al, 2015a). For example, Ploegh et al described a one-pot dual labeling approach for the sequential modification of heterodimeric proteins such as antibodies with different labels at light and heavy chain, respectively, as well as an approach for the sequential C- to -C fusion of two protein of interest (Harmand et al, 2018).…”

Recent Advances in the Discovery and Biosynthetic Study of Eukaryotic RiPP Natural Products

“…A co-crystal structure of butelase 1 with its peptide substrate will help us understand the mechanism of macrocyclization. Due to its high promiscuity and fast kinetics, butelase 1 has been applied in protein labelling [41,42], chemoenzymatic synthesis of bacteriocins [43], generating cyclic peptides with non-native amino acids [44], decorating E. coli cell surfaces [45], making peptide dendrimers [46], and preparing C-to-C fusion proteins [47].…”