Chapter 6 Using Expressed Protein Ligation to Probe the Substrate Specificity of Lantibiotic Synthetases

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

The lantibiotics are a class of ribosomally synthesized and posttranslationally modified peptide antibiotics containing the thioether cross-links lanthionine (Lan) and 3-methyllanthionine (MeLan) and typically also the dehydroamino acids dehydroalanine (Dha) and (Z)-dehydrobutyrine (Dhb). The modifications are formed by dehydration of Ser/Thr residues to produce the Dha and Dhb structures, and subsequent conjugate additions of Cys residues onto the unsaturated amino acids to form thioether rings (Lan and MeLan). Because of their ribosomal origin, investigations of the substrate specificity of lantibiotic synthetases have typically focused on site-directed mutagenesis. With the in vitro reconstitution of lacticin 481 synthetase, its substrate specificity was explored in much greater detail by the incorporation of a series of nonproteinogenic Ser, Thr, and Cys analogues into a truncated prelacticin peptide LctA using a combination of solid-phase peptide synthesis (SPPS) and expressed protein ligation (EPL). The strategy described can be used for the growing number of ribosomally synthesized and posttranslationally modified natural products such as the microcins and patellamides.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
EditorsTom Muir, John Abelson
Pages117-134
Number of pages18
DOIs
StatePublished - Jul 24 2009

Publication series

NameMethods in Enzymology
Volume462
ISSN (Print)0076-6879

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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    Zhang, X., & van der Donk, W. A. (2009). Chapter 6 Using Expressed Protein Ligation to Probe the Substrate Specificity of Lantibiotic Synthetases. In T. Muir, & J. Abelson (Eds.), Methods in Enzymology (pp. 117-134). (Methods in Enzymology; Vol. 462). https://doi.org/10.1016/S0076-6879(09)62006-1