Change search
ReferencesLink to record
Permanent link

Direct link
Isolation and preparation of bacterial cell walls for compositional analysis by Ultra Performance Liquid Chromatography
Umeå University, Faculty of Medicine, Molecular Infection Medicine Sweden (MIMS). Umeå University, Faculty of Medicine, Department of Molecular Biology (Faculty of Medicine).
2014 (English)In: Journal of Visualized Experiments, ISSN 1940-087X, no 83, UNSP e51183- p.Article in journal (Refereed) Published
Abstract [en]

The bacterial cell wall is critical for the determination of cell shape during growth and division, and maintains the mechanical integrity of cells in the face of turgor pressures several atmospheres in magnitude. Across the diverse shapes and sizes of the bacterial kingdom, the cell wall is composed of peptidoglycan, a macromolecular network of sugar strands crosslinked by short peptides. Peptidoglycan's central importance to bacterial physiology underlies its use as an antibiotic target and has motivated genetic, structural, and cell biological studies of how it is robustly assembled during growth and division. Nonetheless, extensive investigations are still required to fully characterize the key enzymatic activities in peptidoglycan synthesis and the chemical composition of bacterial cell walls. High Performance Liquid Chromatography (HPLC) is a powerful analytical method for quantifying differences in the chemical composition of the walls of bacteria grown under a variety of environmental and genetic conditions, but its throughput is often limited. Here, we present a straightforward procedure for the isolation and preparation of bacterial cell walls for biological analyses of peptidoglycan via HPLC and Ultra Performance Liquid Chromatography (UPLC), an extension of HPLC that utilizes pumps to deliver ultra-high pressures of up to 15,000 psi, compared with 6,000 psi for HPLC. In combination with the preparation of bacterial cell walls presented here, the low-volume sample injectors, detectors with high sampling rates, smaller sample volumes, and shorter run times of UPLC will enable high resolution and throughput for novel discoveries of peptidoglycan composition and fundamental bacterial cell biology in most biological laboratories with access to an ultracentrifuge and UPLC.

Place, publisher, year, edition, pages
JoVE , 2014. no 83, UNSP e51183- p.
Keyword [en]
Chemistry, Issue 83, peptidoglycan, bacterial cell wall, ultra-performance liquid chromatography, gh-performance liquid chromatography, cell shape, morphogenesis, LAROSA EJ, 1985, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF
National Category
Microbiology in the medical area Medical Laboratory and Measurements Technologies
URN: urn:nbn:se:umu:diva-100312DOI: 10.3791/51183ISI: 000348513500058PubMedID: 24457605OAI: diva2:792194
Available from: 2015-03-03 Created: 2015-02-27 Last updated: 2015-03-03Bibliographically approved

Open Access in DiVA

No full text

Other links

Publisher's full textPubMed

Search in DiVA

By author/editor
Cava, Felipe
By organisation
Molecular Infection Medicine Sweden (MIMS)Department of Molecular Biology (Faculty of Medicine)
In the same journal
Journal of Visualized Experiments
Microbiology in the medical areaMedical Laboratory and Measurements Technologies

Search outside of DiVA

GoogleGoogle Scholar
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Altmetric score

Total: 35 hits
ReferencesLink to record
Permanent link

Direct link