Journal
BIO-PROTOCOL
Volume 8, Issue 20, Pages -Publisher
BIO-PROTOCOL
DOI: 10.21769/BioProtoc.3063
Keywords
Membrane fluidity; Membrane domains; Laurdan GP; Regions of increased fluidity; RIFs; Bacillus subtilis; Cytoplasmic membrane; Antibiotic mode of action
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Funding
- Netherlands Organization for Scientific Research (NWO) [STW-Vici 12128]
- Amsterdam Infection and Immunity Institute
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Membrane fluidity is a key parameter of bacterial membranes that undergoes quick adaptation in response to environmental challenges and has recently emerged as an important factor in the antibacterial mechanism of membrane-targeting antibiotics. The specific level of membrane fluidity is not uniform across the bacterial cell membrane. Rather, specialized microdomains associated with different cellular functions can exhibit fluidity values that significantly deviate from the average. Assessing changes in the overall membrane fluidity and formation of membrane microdomains is therefore pivotal to understand both the functional organization of the bacterial cell membrane as well as antibiotic mechanisms. Here we describe how two fluorescent membrane dyes, laurdan and DiIC12, can be employed to assess membrane fluidity in living bacteria. We focus on Bacillus subtilis, since this organism has been relatively well-studied with respect to membrane domains. However, we also describe how these assays can be adapted for other bacteria such as Staphylococcus aureus and Streptococcus pneumoniae.
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