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Bioorthogonal Labeling and Click-Chemistry-Based Visualization of the Tannerella forsythia Cell Wall

  • Stephen N. Hyland
  • , Sreedevi Chinthamani
  • , Sushanta Ratna
  • , Kimberly A. Wodzanowski
  • , Liam Michael D. Sandles
  • , Kiyonobu Honma
  • , Catherine Leimkuhler-Grimes
  • , Ashu Sharma
  • University of Delaware
  • SUNY Buffalo

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

1 Scopus citations

Abstract

The objective of this chapter is to provide a detailed protocol for the peptidoglycan (cell wall) labeling of the periodontal pathogen Tannerella forsythia and the development of a laboratory-safe Escherichia coli strain utilizing the N-acetylmuramic acid recycling enzymes AmgK, N-acetylmuramate/N-acetylglucosamine kinase, and MurU, N-acetylmuramate alpha-1-phosphate uridylyltransferase, from T. forsythia. The procedure involves bioorthogonal labeling of bacterial cells with an azido-modified analog of the amino sugar, N-acetylmuramic acid, through “click chemistry” with a fluorescent dye. The protocol is suitable for the generation of fluorescently labeled peptidoglycan molecules for applications in the study of bacterial and peptidoglycan trafficking in the host cells and cell wall recycling in complex microbiomes.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages1-16
Number of pages16
DOIs
StatePublished - 2024

Publication series

NameMethods in Molecular Biology
Volume2727
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Bioorthogonal
  • Cell wall recycling
  • Click chemistry
  • Mass spectrometry
  • Microscopy
  • Peptidoglycan
  • Tannerella forsythia

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