A method for characterizing the thermal stability and antimicrobial binding to Lipopolysaccharides of Gram-negative isogenic mutant strains

Belén Navarro, Mackarenna Alarcón, Maricarmen Osees, Felipe Gómez-Alvear, Romina V. Sepúlveda, Jaime Huerta, María Cecilia Opazo, Daniel Aguayo

Research output: Contribution to journalArticlepeer-review

Abstract

Isothermal Titration Calorimetry (ITC) is widely employed to assess antimicrobial affinity for lipopolysaccharide (LPS); nevertheless, experiments are usually limited to commercially available-LPS chemotypes. Herein we show a method that uses Differential Scanning Calorimetry (DSC) to characterize homogeneity artificial vesicles of LPS (LPS-V) extracted from isogenic mutant bacterial strains before analyzing the antimicrobial binding by ITC. This method allows us to characterize the differences in the Polymyxin-B binding and gel to crystalline liquid (β↔α) phase profiles of LPS-V made of LPS extracted from Escherichia coli isogenic mutant strains for the LPS biosynthesis pathway, allowing us to obtain the comparable data required for new antimicrobial discovery. A method for: • Obtaining LPS vesicles from isogenic mutant bacterial strains. • Characterize artificial LPS vesicles homogeneity. • Characterize antimicrobial binding to LPS.

Original languageEnglish
Article number101474
JournalMethodsX
Volume8
DOIs
Publication statusPublished - Jan 2021

Keywords

  • Antimicrobial binding
  • Calorimetric methods to study Lipopolysaccharide-vesicles
  • Differential Scanning Calorimetry (ITC)
  • Isothermal Titration Calorimetry (ITC)
  • Lipopolysaccharides

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Medical Laboratory Technology

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