Issue 12, 2024

Sequence-function space of radical SAM cyclophane synthases reveal conserved active site residues that influence substrate specificity

Abstract

Radical SAM cyclophane synthases catalyze C–C, C–N, and C–O crosslinking reactions in the biosynthesis of bioactive peptide natural products. Here, we studied an uncharacterized rSAM enzyme, HtkB from Pandoraea sp., and found this enzyme to catalyze the formation of a HisC2-to-LysCβ crosslink. We used a combination of ColabFold and mutagenesis studies to show that residues D214 in HtkB and H204 in HaaB (another cyclophane synthase) are important for substrate specificity. Mutation of these residues changes the specificity and lowers substrate recognition on the wild-type motifs. This result opens opportunities to alter the specificity and promiscuity for rSAM peptide modifying enzymes.

Graphical abstract: Sequence-function space of radical SAM cyclophane synthases reveal conserved active site residues that influence substrate specificity

Supplementary files

Article information

Article type
Communication
Submitted
24 Sep 2024
Accepted
18 Oct 2024
First published
23 Oct 2024
This article is Open Access
Creative Commons BY-NC license

RSC Chem. Biol., 2024,5, 1195-1200

Sequence-function space of radical SAM cyclophane synthases reveal conserved active site residues that influence substrate specificity

C. Phan and B. I. Morinaka, RSC Chem. Biol., 2024, 5, 1195 DOI: 10.1039/D4CB00227J

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