Butelase: Linkage‐specific Ligase

Abstract

Site‐specific ligation is enormously useful to enable selective bonding of chemicals, polymers, peptides, and proteins to form new compounds. Recent advances in chemistry, biochemistry and molecular biology have provided novel methods to staple peptide and non‐peptide bonds under physiological conditions. An example is the family of linkage‐specific, peptide asparaginyl‐specific ligases (PAL). A prototypic member of PAL is butelase 1 isolated from the butterfly pea (Bunga Telang). Butelase 1, catalyzing the reverse reaction of an asparaginyl endopeptidase, is a stand‐alone and ATP‐independent ligase [1]. Importantly, butelase 1 is C‐terminus‐specific for Asn/Asp (Asx), traceless, and accepts a tripeptide Asx‐His‐Val with the dipeptide His‐Val as the leaving group. Butelase 1 accepts most N‐terminal amino acids with D‐or L‐configuration. It exhibits unmatched kinetics with catalytic efficiencies of up to 1,340,000 M−1 s−1 and is >10,000 times faster than other known ligases. Our recently published work showed that butelase 1 is useful for both intra‐ and intermolecular ligation. It efficiently cyclizes or ligates various peptides and proteins ranging in size from 8 to >300 amino acids. Thus, the high catalytic efficiency and broad substrate specificity of butelase 1 could augment new applications, both in in vitro and in vivo systems for basic and translational research. Here, I will present our latest results on Asx‐specific ligases [2–6], their applications, and live‐cell labeling to explore new frontiers in biochemical, medical and material sciences.Support or Funding InformationThis research was supported in part by Nanyang Technological University Internal Funding ‐ Synzymes and Natural Products (SYNC) and the AcRF Tier 3 funding (MOE2016‐T3‐1‐003).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.