9IY4

Iterative acetyltransferase on lasso peptides from Actinomycetes in complex with AcCoA

Summary for 9IY4

• Entry DOI: 10.2210/pdb9iy4/pdb
• EMDB information: 60984
• Descriptor: GCN5-related N-acetyltransferase, ACETYL COENZYME *A (2 entities in total)
• Functional Keywords: gcn5-related n-acetyltransferases, actinomycetes, accoa, transferase
• Biological source: Actinosynnema mirum (strain ATCC 29888 / DSM 43827 / JCM 3225 / NBRC 14064 / NCIMB 13271 / NRRL B-12336 / IMRU 3971 / 101)
• Total number of polymer chains: 1
• Total formula weight: 22485.74

Authors

Wu, S.,Xiong, J.,Lei, D.,Dong, S. (deposition date: 2024-07-29, release date: 2025-08-13, Last modification date: 2026-04-01)

Primary citation

Xiong, J.,Wu, S.,Liang, Z.Q.,Fang, S.,Tao, F.Y.,Gong, X.T.,Wu, X.,Wu, Q.,Cui, J.J.,Gao, K.,Hoi, K.K.,Peng, Y.,Luo, S.,Lei, D.,Dong, S.H.
Iterative acylation on mature lasso peptides by widespread acetyltransferases.
Nat.Chem.Biol., 2026

PubMed Abstract: The biosynthesis of ribosomally synthesized and posttranslationally modified peptides (RiPPs) leverages iterative catalysis to enhance structural and biological diversity. Traditionally, iterative enzymes install posttranslational modifications on linear peptides, rather than mature RiPPs with intricate three-dimensional structures, which require complex changes in substrate binding. Here we present a prolific class of GCN5-related N-acetyltransferases (GNATs) that iteratively and consecutively acylate two Lys residues within the loop and ring motifs of lasso peptides, diverging from the typical iterative modification of linear peptides. Utilizing high-resolution cryogenic-electron microscopy and enzymatic reconstitution, we define the lasso peptide-binding pocket of IatT and pinpoint key residues that distinguish its two distinct acetylation steps. Structure-based engineering of IatT's acetyl-recognition site expands the cavity to accommodate longer-chain acyl groups, enabling the creation of lipolasso peptides, a class of ribosomal lipopeptide. This engineering strategy can be applied to any RiPP biosynthetic gene cluster encoding GNAT, facilitating the efficient diversification of ribosomal lipopeptides.

PubMed: 41826761
DOI: 10.1038/s41589-026-02149-6

Experimental method

ELECTRON MICROSCOPY (2 Å)