Bulletin of the American Physical Society
APS March Meeting 2023
Volume 68, Number 3
Las Vegas, Nevada (March 5-10)
Virtual (March 20-22); Time Zone: Pacific Time
Session B10: Quantitative Cell Physiology I - Shape and Size
11:30 AM–2:30 PM,
Monday, March 6, 2023
Room: Room 202
Sponsoring
Unit:
DBIO
Chair: Suckjoon Jun, University of California, San Diego
Abstract: B10.00011 : Mapping nanostructural changes in E.coli Peptidoglycan*
1:30 PM–1:42 PM
Presenter:
Abimbola F Olulana
(Department of Physics and Astronomy, University of Sheffield)
Authors:
Abimbola F Olulana
(Department of Physics and Astronomy, University of Sheffield)
Jacob Biboy
(Centre for Bacterial Cell Biology, Biosciences Institute, Newcastle University, UK)
Oliver Meacock
(Department of Physics and Astronomy, University of Sheffield, UK)
Laia Pasquina-Lemonche
(Department of Physics and Astronomy, University of Sheffield, UK)
William M Durham
(Department of Physics and Astronomy, University of Sheffield, UK)
Simon J Foster
(Department of Molecular Biology and Biotechnology, University of Sheffield, UK)
Waldemar Vollmer
(Centre for Bacterial Cell Biology, Biosciences Institute, Newcastle University, UK)
Jamie K Hobbs
(Department of Physics and Astronomy, University of Sheffield, Sheffield, UK)
To quantify the associated nanoscale directionality and orientation, we developed a MATLAB script that uses ridge detection to automatically select strands in the AFM image, quantify its orientation, and output an orientation color map and angular distribution plot. In addition, we developed an image segmentation pipeline—a combination of Otsu thresholding and particle analysis to measure gaps, holes, and breaks in the PG network. Using the aforementioned approaches, our results reveal unprecedented detail of nanometric molecular organizations with location-dependent orientation across the PG and the distribution of holes and breaks as a function of antibiotic treatment times. The increase in the frequency and size of breaks suggest the progression of death due to antibiotic-induced enzyme inactivation. Our findings reveal the remarkable application of high-resolution AFM in deciphering the bacterial cell wall; bringing us closer to the understanding of how antibiotics affect PG molecular organization and in turn, regulates its function & stability.
References
1. Vollmer, W., Blanot, D. & De Pedro, M. A. Peptidoglycan structure and architecture. FEMS Microbiol. Rev. 32, 149–167 (2008).
2. Vollmer, W. & Bertsche, U. Murein (peptidoglycan) structure, architecture and biosynthesis in Escherichia coli. Biochim. Biophys. Acta - Biomembr. 1778, 1714–1734 (2008).
*This work is fully funded by Engineering and Physical Sciences Research Council (EPSRC, EP/T002778/1).
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