Abstract
Lipopolysaccharide (LPS) on Gram-negative bacterial outer membranes is the first target for antimicrobial agents, due to their spatial proximity to outer environments of microorganisms. To understand the molecular nature and their interaction with antimicrobial agents, establishing a model LPS structure is of key importance. Here, we describe procedures for following LPS layer attachment to a solid surface and provide protocols for measuring bacterial membrane morphology after adding antibiotics. Using atomic force microscopy (AFM), we show methods to characterize the effects of antibiotic polymyxin B to the LPS layers at the nanoscale.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Oh YJ, Cui Y, Kim H, Li Y, Hinterdorfer P, Park S (2012) Characterization of curli A production on living bacterial surfaces by scanning probe microscopy. Biophys J 103(8):1666–1671. https://doi.org/10.1016/j.bpj.2012.09.004
Vadillo-Rodriguez V, Beveridge TJ, Dutcher JR (2008) Surface viscoelasticity of individual gram-negative bacterial cells measured using atomic force microscopy. J Bacteriol 190(12):4225–4232. https://doi.org/10.1128/jb.00132-08
Sperandeo P, Martorana AM, Polissi A (2017) Lipopolysaccharide biogenesis and transport at the outer membrane of Gram-negative bacteria. Biochimica et Biophysica Acta (BBA) – Mol Cell Biol Lipids 1862(11):1451–1460. https://doi.org/10.1016/j.bbalip.2016.10.006
Oh YJ, Plochberger B, Rechberger M, Hinterdorfer P (2017) Characterizing the effect of polymyxin B antibiotics to lipopolysaccharide on Escherichia coli surface using atomic force microscopy. J Mol Recognit 30(6):e2605. https://doi.org/10.1002/jmr.2605
Cui Y, Oh YJ, Lim J, Youn M, Lee I, Pak HK, Park W, Jo W, Park S (2012) AFM study of the differential inhibitory effects of the green tea polyphenol (−)-epigallocatechin-3-gallate (EGCG) against gram-positive and gram-negative bacteria. Food Microbiol 29(1):80–87. https://doi.org/10.1016/j.fm.2011.08.019
Dufrêne YF (2002) Atomic force microscopy, a powerful tool in microbiology. J Bacteriol 184(19):5205–5213. https://doi.org/10.1128/jb.184.19.5205-5213.2002
Dufrêne YF (2014) Atomic force microscopy in microbiology: new structural and functional insights into the microbial cell surface. mBio 5(4):e01363–e01314. https://doi.org/10.1128/mBio.01363-14
Dupres V, Alsteens D, Andre G, Dufrêne YF (2010) Microbial nanoscopy: a closer look at microbial cell surfaces. Trend Microbiol 18(9):397–405. https://doi.org/10.1016/j.tim.2010.06.004
Rico F, Su C, Scheuring S (2011) Mechanical mapping of single membrane proteins at submolecular resolution. Nano Lett 11(9):3983–3986. https://doi.org/10.1021/nl202351t
Mortensen NP, Fowlkes JD, Sullivan CJ, Allison DP, Larsen NB, Molin S, Doktycz MJ (2009) Effects of Colistin on surface ultrastructure and nanomechanics of Pseudomonas aeruginosa cells. Langmuir 25(6):3728–3733. https://doi.org/10.1021/la803898g
Formosa C, Grare M, Jauvert E, Coutable A, Regnouf-de-Vains JB, Mourer M, Duval RE, Dague E (2012) Nanoscale analysis of the effects of antibiotics and CX1 on a Pseudomonas aeruginosa multidrug-resistant strain. Sci Rep 2(1):575. https://doi.org/10.1038/srep00575
Oh YJ, Hinterdorfer P (2019) Investigation of bacterial Curli production and adhesion using AFM. Method Mol Biol (Clifton, NJ) 1886:221–231. https://doi.org/10.1007/978-1-4939-8894-5_12
Artelsmair H, Kienberger F, Tinazli A, Schlapak R, Zhu R, Preiner J, Wruss J, Kastner M, Saucedo-Zeni N, Hoelzl M, Rankl C, Baumgartner W, Howorka S, Blaas D, Gruber HJ, Tampé R, Hinterdorfer P (2008) Atomic force microscopy-derived nanoscale chip for the detection of human pathogenic viruses. Small 4(6):847–854. https://doi.org/10.1002/smll.200700691
Acknowledgment
Y.O. acknowledges the financial support from the Austrian Science Fund FWF project V584-BBL and the Austrian National Foundation for Research, Technology, and Development and Research Department of the State of Upper Austria.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Oh, Y.J. (2022). Use of Atomic Force Microscopy to Characterize LPS Perturbations. In: Sperandeo, P. (eds) Lipopolysaccharide Transport. Methods in Molecular Biology, vol 2548. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2581-1_17
Download citation
DOI: https://doi.org/10.1007/978-1-0716-2581-1_17
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2580-4
Online ISBN: 978-1-0716-2581-1
eBook Packages: Springer Protocols