High-throughput, Highly Sensitive Analyses of Bacterial Morphogenesis Using Ultra Performance Liquid Chromatography.

The Journal of Biological Chemistry
Samantha M DesmaraisKerwyn Casey Huang

Abstract

The bacterial cell wall is a network of glycan strands cross-linked by short peptides (peptidoglycan); it is responsible for the mechanical integrity of the cell and shape determination. Liquid chromatography can be used to measure the abundance of the muropeptide subunits composing the cell wall. Characteristics such as the degree of cross-linking and average glycan strand length are known to vary across species. However, a systematic comparison among strains of a given species has yet to be undertaken, making it difficult to assess the origins of variability in peptidoglycan composition. We present a protocol for muropeptide analysis using ultra performance liquid chromatography (UPLC) and demonstrate that UPLC achieves resolution comparable with that of HPLC while requiring orders of magnitude less injection volume and a fraction of the elution time. We also developed a software platform to automate the identification and quantification of chromatographic peaks, which we demonstrate has improved accuracy relative to other software. This combined experimental and computational methodology revealed that peptidoglycan composition was approximately maintained across strains from three Gram-negative species despite taxonomical an...Continue Reading

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Citations

Apr 28, 2016·Biophysical Journal·Joris M H GoudsmitsAndrew Robinson
Aug 16, 2016·Analytical and Bioanalytical Chemistry·Marshall BernStéphane Mesnage
Jan 14, 2017·Cell·Thomas M BartlettZemer Gitai
Jul 10, 2019·Molecular Microbiology·Anna I WeaverTobias Dörr
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Aug 29, 2019·ELife·Nikola OjkicShiladitya Banerjee
Jul 20, 2018·Nature·Enrique R RojasKerwyn Casey Huang
Nov 17, 2020·Frontiers in Cellular and Infection Microbiology·Ann-Britt Schäfer, Michaela Wenzel
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Nov 5, 2019·Bio-protocol·Gabriele Stankeviciute, Eric A Klein
Jan 21, 2021·EcoSal Plus·Shambhavi GardeManjula Reddy

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