Structural investigation of lanthipeptides involved in substrate controlled pathways and methods for leader peptide removal
type: CreativeWork
description: Ribosomally synthesized and post-translationally modified peptides are a family of natural products with diverse activities, with lanthipeptides as one of the most prominent classes. The diverse ring topology of lanthipeptides is formed by four classes of enzymes with substrate tolerance that holds potential for development of biotechnological techniques for macrocyclizations that may increase the utility of peptide therapeutics. The general biosynthetic scheme of a leader peptide that plays a role in the processing of the core peptide by biosynthetic enzymes is powerful as such pathways could potentially be used to make products that are very different from the natural product. This strategy would be most successful when the structural characterization and mechanism of lanthionine synthetases is fully understood. However, a difficult obstacle for high throughput studies of these natural products is removal of the leader peptide. Chapter 2 discusses the development of a general method using unnatural amino acid incorporation to replace the peptide bond between the leader peptide and the core peptide with an ester bond, which can be easily hydrolyzed in basic environments. This proof-of-concept method was site selective for the tested substrates setting the stage for further development and optimization for increasing the yields of peptides produced by this route. Chapter 3 extends this theme through the identification of a general protease for the removal of double glycine motif-leader peptides, used in a subset of RiPPs which had escaped the development of a general method for leader peptide removal. The cognate proteases in lanthipeptide biosynthetic machineries are often membrane associated proteins that suffer from a decrease in catalytic activity when taken out of their cellular environment and/or that are not active at if segregated from the membrane-bound domains. The structural characterization of an isolated protease domain of a LanT-type transporter from L…
abstract: Ribosomally synthesized and post-translationally modified peptides are a family of natural products with diverse activities, with lanthipeptides as one of the most prominent classes. The diverse ring topology of lanthipeptides is formed by four classes of enzymes with substrate tolerance that holds potential for development of biotechnological techniques for macrocyclizations that may increase the utility of peptide therapeutics. The general biosynthetic scheme of a leader peptide that plays a role in the processing of the core peptide by biosynthetic enzymes is powerful as such pathways could potentially be used to make products that are very different from the natural product. This strategy would be most successful when the structural characterization and mechanism of lanthionine synthetases is fully understood. However, a difficult obstacle for high throughput studies of these natural products is removal of the leader peptide. Chapter 2 discusses the development of a general method using unnatural amino acid incorporation to replace the peptide bond between the leader peptide and the core peptide with an ester bond, which can be easily hydrolyzed in basic environments. This proof-of-concept method was site selective for the tested substrates setting the stage for further development and optimization for increasing the yields of peptides produced by this route. Chapter 3 extends this theme through the identification of a general protease for the removal of double glycine motif-leader peptides, used in a subset of RiPPs which had escaped the development of a general method for leader peptide removal. The cognate proteases in lanthipeptide biosynthetic machineries are often membrane associated proteins that suffer from a decrease in catalytic activity when taken out of their cellular environment and/or that are not active at if segregated from the membrane-bound domains. The structural characterization of an isolated protease domain of a LanT-type transporter from L…
author: Bobeica, Silvia Cristina
language: en
keywords: lanthipeptide, RiPP, ribosomally synthesized and post-translationally modified peptides, thioether, lanthionine, methyllanthionine, prochlorosin, cytolysin
date published: 2020-10-07T20:59:35Z
date modified: 2026-07-22T22:24:48Z
provider: University of Illinois - Urbana-Champaign
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: University of Illinois at Urbana-Champaign

PropertyValue
property ID: Handle
value: http://hdl.handle.net/2142/108443