A lanthipeptide library used to identify a protein-protein interaction inhibitor

Xiao Yang; Katherine R Lennard; Chang He; Mark C Walker; Andrew T Ball; Cyrielle Doigneaux; Ali Tavassoli; Wilfred A van der Donk

doi:10.1038/s41589-018-0008-5

A lanthipeptide library used to identify a protein-protein interaction inhibitor

Nat Chem Biol. 2018 Apr;14(4):375-380. doi: 10.1038/s41589-018-0008-5. Epub 2018 Mar 5.

Authors

Xiao Yang¹, Katherine R Lennard², Chang He¹, Mark C Walker¹, Andrew T Ball², Cyrielle Doigneaux², Ali Tavassoli³, Wilfred A van der Donk⁴

Affiliations

¹ Department of Chemistry and Howard Hughes Medical Institute, University of Illinois at Urbana-Champaign, Urbana, IL, USA.
² Department of Chemistry and Institute for Life Sciences, University of Southampton, Southampton, UK.
³ Department of Chemistry and Institute for Life Sciences, University of Southampton, Southampton, UK. a.tavassoli@soton.ac.uk.
⁴ Department of Chemistry and Howard Hughes Medical Institute, University of Illinois at Urbana-Champaign, Urbana, IL, USA. vddonk@illinois.edu.

Abstract

In this article we describe the production and screening of a genetically encoded library of 10⁶ lanthipeptides in Escherichia coli using the substrate-tolerant lanthipeptide synthetase ProcM. This plasmid-encoded library was combined with a bacterial reverse two-hybrid system for the interaction of the HIV p6 protein with the UEV domain of the human TSG101 protein, which is a critical protein-protein interaction for HIV budding from infected cells. Using this approach, we identified an inhibitor of this interaction from the lanthipeptide library, whose activity was verified in vitro and in cell-based virus-like particle-budding assays. Given the variety of lanthipeptide backbone scaffolds that may be produced with ProcM, this method may be used for the generation of genetically encoded libraries of natural product-like lanthipeptides containing substantial structural diversity. Such libraries may be combined with any cell-based assay to identify lanthipeptides with new biological activities.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Crystallography, X-Ray
DNA-Binding Proteins / chemistry*
Endosomal Sorting Complexes Required for Transport / chemistry*
ErbB Receptors / metabolism
Escherichia coli / metabolism*
Escherichia coli Proteins / chemistry
Ethylmaleimide / chemistry
Gene Library
HEK293 Cells
HIV
HeLa Cells
Humans
Inhibitory Concentration 50
Iodoacetamide / chemistry
Peptide Hydrolases / chemistry
Peptide Synthases / chemistry*
Peptides / chemistry*
Plasmids
Protein Domains
Protein Interaction Mapping
Substrate Specificity
Transcription Factors / chemistry*
gag Gene Products, Human Immunodeficiency Virus / chemistry*

Substances

DNA-Binding Proteins
Endosomal Sorting Complexes Required for Transport
Escherichia coli Proteins
Peptides
Transcription Factors
Tsg101 protein
gag Gene Products, Human Immunodeficiency Virus
p6 gag protein, Human immunodeficiency virus 1
EGFR protein, human
ErbB Receptors
Peptide Hydrolases
Peptide Synthases
Ethylmaleimide
Iodoacetamide

Abstract

Publication types

MeSH terms

Substances

Grants and funding