Characterization of a dual function macrocyclase enables design and use of efficient macrocyclization substrates.

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.

journal_name

Nat Commun

journal_title

Nature communications

authors

Czekster CM,Ludewig H,McMahon SA,Naismith JH

doi

10.1038/s41467-017-00862-4

subject

Has Abstract

pub_date

2017-10-19 00:00:00

pages

1045

issue

1

issn

2041-1723

pii

10.1038/s41467-017-00862-4

journal_volume

8

pub_type

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