Molecular Basis for the Broad Substrate Selectivity of a Peptide Prenyltransferase

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2016 Nov 23

PMID 27872314

Citations 28

Authors

Yue Hao

Elizabeth Pierce

Daniel Roe

Maho Morita

John A McIntosh

Vinayak Agarwal

Thomas E Cheatham 3rd

Eric W Schmidt

Satish K Nair

Affiliations

Soon will be listed here.

Abstract

The cyanobactin prenyltransferases catalyze a series of known or unprecedented reactions on millions of different substrates, with no easily observable recognition motif and exquisite regioselectivity. Here we define the basis of broad substrate tolerance for the otherwise uncharacterized TruF family. We determined the structures of the Tyr-prenylating enzyme PagF, in complex with an isoprenoid donor analog and a panel of linear and macrocyclic peptide substrates. Unexpectedly, the structures reveal a truncated barrel fold, wherein binding of large peptide substrates is necessary to complete a solvent-exposed hydrophobic pocket to form the catalytically competent active site. Kinetic, mutational, chemical, and computational analyses revealed the structural basis of selectivity, showing a small motif within peptide substrates that is sufficient for recognition by the enzyme. Attaching this 2-residue motif to two random peptides results in their isoprenylation by PagF, demonstrating utility as a general biocatalytic platform for modifications on any peptide substrate.

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