TY - JOUR
T1 - Pandonodin
T2 - A Proteobacterial Lasso Peptide with an Exceptionally Long C-Terminal Tail
AU - Cheung-Lee, Wai Ling
AU - Cao, Li
AU - Link, A. James
N1 - Funding Information:
The atomic coordinates for the pandonodin NMR structure have been deposited in the PDB (accession number 6Q1X ) and the BMRB (accession number 30651). This study was supported in part by National Institutes of Health Grant GM107036 to A.J.L. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. L.C. was supported by the National Science Foundation Graduate Research Fellowship Program under Grant DGE-1656466. W.L.C.-L. was supported in part by a Dodds Fellowship from Princeton University. The authors declare no competing financial interest.
Publisher Copyright:
Copyright © 2019 American Chemical Society.
PY - 2019/12/20
Y1 - 2019/12/20
N2 - 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.
AB - 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.
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U2 - 10.1021/acschembio.9b00676
DO - 10.1021/acschembio.9b00676
M3 - Article
C2 - 31742991
AN - SCOPUS:85076240608
SN - 1554-8929
VL - 14
SP - 2783
EP - 2792
JO - ACS chemical biology
JF - ACS chemical biology
IS - 12
ER -