Molecular basis for the broad substrate selectivity of a peptide prenyltransferase

Yue Hao, Elizabeth Pierce, Daniel Roe, Maho Morita, John A. McIntosh, Vinayak Agarwal, Thomas E. Cheatham, Eric W. Schmidt, Satish K. Nair

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish (US)
Pages (from-to)14037-14042
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume113
Issue number49
DOIs
StatePublished - Dec 6 2016

Keywords

  • Biosynthesis
  • Crystallography
  • Prenylation
  • RiPP

ASJC Scopus subject areas

  • General

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