Czekster_2017_Nat.Commun_8_1045

Reference

Title : Characterization of a dual function macrocyclase enables design and use of efficient macrocyclization substrates - Czekster_2017_Nat.Commun_8_1045
Author(s) : Czekster CM , Ludewig H , McMahon SA , Naismith JH
Ref : Nat Commun , 8 :1045 , 2017
Abstract :

Peptide macrocycles are promising therapeutic molecules because they are protease resistant, structurally rigid, membrane permeable, and capable of modulating protein-protein interactions. Here, we report the characterization of the dual function macrocyclase-peptidase enzyme involved in the biosynthesis of the highly toxic amanitin toxin family of macrocycles. The enzyme first removes 10 residues from the N-terminus of a 35-residue substrate. Conformational trapping of the 25 amino-acid peptide forces the enzyme to release this intermediate rather than proceed to macrocyclization. The enzyme rebinds the 25 amino-acid peptide in a different conformation and catalyzes macrocyclization of the N-terminal eight residues. Structures of the enzyme bound to both substrates and biophysical analysis characterize the different binding modes rationalizing the mechanism. Using these insights simpler substrates with only five C-terminal residues were designed, allowing the enzyme to be more effectively exploited in biotechnology.

PubMedSearch : Czekster_2017_Nat.Commun_8_1045
PubMedID: 29051530
Gene_locus related to this paper: 9agar-h2e7q8

Related information

Substrate Alpha-amanitin-proprotein
Gene_locus Alpha-amanitin-proprotein    9agar-h2e7q8
Structure Alpha-amanitin-proprotein    9agar-h2e7q8    5N4B    5N4C    5N4D    5N4E    5N4F

Citations formats

Czekster CM, Ludewig H, McMahon SA, Naismith JH (2017)
Characterization of a dual function macrocyclase enables design and use of efficient macrocyclization substrates
Nat Commun 8 :1045

Czekster CM, Ludewig H, McMahon SA, Naismith JH (2017)
Nat Commun 8 :1045