5O3W

Structural characterization of the fast and promiscuous macrocyclase from plant - PCY1-S562A bound to Presegetalin A1

Summary for 5O3W

• Entry DOI: 10.2210/pdb5o3w/pdb
• Descriptor: Peptide cyclase 1, Presegetalin A1, MAGNESIUM ION, ... (5 entities in total)
• Functional Keywords: segetalin biosynthesis, prolyl oligopeptidase, macrocyclase, peptidase, beta-propeller, closed form, hydrolase
• Biological source: Vaccaria hispanica; More
• Total number of polymer chains: 8
• Total formula weight: 343793.43

Authors

Ludewig, H.,Czekster, C.M.,Bent, A.F.,Naismith, J.H. (deposition date: 2017-05-25, release date: 2018-02-07, Last modification date: 2024-05-01)

Primary citation

Ludewig, H.,Czekster, C.M.,Oueis, E.,Munday, E.S.,Arshad, M.,Synowsky, S.A.,Bent, A.F.,Naismith, J.H.
Characterization of the Fast and Promiscuous Macrocyclase from Plant PCY1 Enables the Use of Simple Substrates.
ACS Chem. Biol., 13:801-811, 2018

PubMed Abstract: Cyclic ribosomally derived peptides possess diverse bioactivities and are currently of major interest in drug development. However, it can be chemically challenging to synthesize these molecules, hindering the diversification and testing of cyclic peptide leads. Enzymes used in vitro offer a solution to this; however peptide macrocyclization remains the bottleneck. PCY1, involved in the biosynthesis of plant orbitides, belongs to the class of prolyl oligopeptidases and natively displays substrate promiscuity. PCY1 is a promising candidate for in vitro utilization, but its substrates require an 11 to 16 residue C-terminal recognition tail. We have characterized PCY1 both kinetically and structurally with multiple substrate complexes revealing the molecular basis of recognition and catalysis. Using these insights, we have identified a three residue C-terminal extension that replaces the natural recognition tail permitting PCY1 to operate on synthetic substrates. We demonstrate that PCY1 can macrocyclize a variety of substrates with this short tail, including unnatural amino acids and nonamino acids, highlighting PCY1's potential in biocatalysis.

PubMed: 29377663
DOI: 10.1021/acschembio.8b00050

Experimental method

X-RAY DIFFRACTION (2 Å)