Pandonodin: A Proteobacterial Lasso Peptide with an Exceptionally Long C-Terminal Tail.

TitlePandonodin: A Proteobacterial Lasso Peptide with an Exceptionally Long C-Terminal Tail.
Publication TypeJournal Article
Year of Publication2019
AuthorsCheung-Lee, WLing, Cao, L, A Link, J
JournalACS Chem Biol
Volume14
Issue12
Pagination2783-2792
Date Published2019 12 20
ISSN1554-8937
KeywordsAmino Acid Sequence, Biophysical Phenomena, Multigene Family, Nuclear Magnetic Resonance, Biomolecular, Peptides, Protein Conformation, Protein Stability, Proteobacteria
Abstract

<p>Lasso peptides are a family of ribosomally synthesized and post-translationally modified peptides (RiPPs) defined by their threaded-ring topology. The N-terminus of the peptide forms an isopeptide bond with an aspartate or glutamate side chain to create a 7-9 amino acid (aa) macrocyclic ring through which the rest of the peptide is threaded. The result is a highly constrained three-dimensional structure. Even though they share a threaded-ring feature, characterized lasso peptides vary greatly in sequence and size, ranging from 14 to 26 aa. Using genome mining, we identified a new lasso peptide gene cluster with a predicted lasso peptide that is 33 aa long. Here we report the heterologous expression of this new peptide, pandonodin, its NMR structure, and its unusual biophysical properties. Pandonodin has a long, proteolytically resistant 18-residue tail of low sequence complexity, which limits its water solubility. Within this tail is a 6 aa disulfide-bonded macrocycle that serves as a steric lock to maintain the lasso structure. This disulfide bond is unusually stable, requiring both heat and high concentrations of reductants for cleavage. Finally, we also show that segments of the C-terminal tail of pandonodin can be replaced with arbitrary sequences, allowing for the construction of pandonodin-protein fusions.</p>

DOI10.1021/acschembio.9b00676
Alternate JournalACS Chem. Biol.
PubMed ID31742991
PubMed Central IDPMC7010350
Grant ListR01 GM107036 / GM / NIGMS NIH HHS / United States