Ceramide-mediation of diffusion in supported lipid bilayers

Hossain, Masroor; Blanchard, G. J.

doi:10.1016/j.chemphyslip.2021.105090

Cited by 2 publications

(3 citation statements)

References 98 publications

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“…An alternative to Laurdan is the fluorescence anisotropy probe 1,6‐diphenyl‐1,3,5‐hexatriene, which delivers comparable results but requires a spectrometer equipped with a polarizer ( 50 ). A dye-independent measure of membrane fluidity is fluorescence recovery after photobleaching, which assesses diffusion of protein fusions or dyes in the membrane ( 51 – 53 ), yet this requires a suitably equipped confocal fluorescence microscope and is thus not accessible to many researchers.…”

Section: Resultsmentioning

confidence: 99%

Dissecting antibiotic effects on the cell envelope using bacterial cytological profiling: a phenotypic analysis starter kit

Schäfer,

Sidarta,

Abdelmesseh Nekhala

et al. 2024

Microbiol Spectr

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Phenotypic analysis assays such as bacterial cytological profiling (BCP) have become increasingly popular for antibiotic mode of action analysis. A plethora of dyes, protein fusions, and reporter strains are available and have been used for this purpose, enabling both rapid mode of action categorization and in-depth analysis of antibiotic mechanisms. However, non-expert researchers may struggle choosing suitable assays and interpreting results. This is a particular problem for antibiotics that have multiple or complex targets, such as the bacterial cell envelope. Here, we set out to curate a minimal set of accessible and affordable phenotypic assays that allow distinction between membrane and cell wall targets, can identify dual-action inhibitors, and can be implemented in most research environments. To this end, we employed BCP, membrane potential, fluidity, and cell wall synthesis assays. To assess specificity and ease of interpretation, we tested three well-characterized and commercially available reference antibiotics: the potassium ionophore valinomycin, the lipid II-binding glycopeptide vancomycin, and the dual-action lantibiotic nisin, which binds lipid II and forms a membrane pore. Based on our experiments, we suggest a minimal set of BCP, a membrane-potentiometric probe, and fluorescent protein fusions to MinD and MreB as basic assay set and recommend complementing these assays with Laurdan-based fluidity measurements and a PliaI reporter fusion, where indicated. We believe that our results can provide guidance for researchers who wish to use phenotypic analysis for mode of action studies but do not possess the specialized equipment or expert knowledge to employ the full breadth of possible techniques. IMPORTANCE Phenotypic analysis assays using specialized fluorescence fusions and dyes have become increasingly popular in antibiotic mode of action analysis. However, it can be difficult to implement these methods due to the need for specialized equipment and/or the complexity of bacterial cell biology and physiology, making the interpretation of results difficult for non-experts. This is especially problematic for compounds that have multiple or pleiotropic effects, such as inhibitors of the bacterial cell envelope. In order to make phenotypic analysis assays accessible to labs, whose primary expertise is not bacterial cell biology, or with limited equipment and resources, a set of simple and broadly accessible assays is needed that is easy to implement, execute, and interpret. Here, we have curated a set of assays and strains that does not need highly specialized equipment, can be performed in most labs, and is straightforward to interpret without knowing the intricacies of bacterial cell biology.

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Section: Resultsmentioning

confidence: 99%

Dissecting antibiotic effects on the cell envelope using bacterial cytological profiling: a phenotypic analysis starter kit

Schäfer,

Sidarta,

Abdelmesseh Nekhala

et al. 2024

Microbiol Spectr

View full text Add to dashboard Cite

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“…Additionally, stress conditions can escalate the secretion of sphingomyelinases, inducing sphingomyelin (SM) to degrade into ceramide. − Elevated ceramide levels in lipid domains are linked to cholesterol depletion . This chemical transition significantly influences the plasma membrane properties, affecting orderliness, topology, and curvature. , Interestingly, ceramides can link together within the plasma membrane, forming gel-like, ceramide-rich platforms . This leads to a profound restructuring of the lipid domain organization as small domains transform into large ceramide-rich signaling platforms …”

Section: The Lipid Landscape At the 5-ht Synapsementioning

confidence: 99%

“…132 This chemical transition significantly influences the plasma membrane properties, affecting orderliness, topology, and curvature. 133,134 Interestingly, ceramides can link together within the plasma membrane, forming gel-like, ceramide-rich platforms. 135 This leads to a profound restructuring of the lipid domain organization as small domains transform into large ceramiderich signaling platforms.…”

Section: ■ Introductionmentioning

confidence: 99%

Serotonin Signaling through Lipid Membranes

Kalinichenko,

Kornhuber,

Sinning

et al. 2024

ACS Chem. Neurosci.

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Serotonin (5-HT) is a vital modulatory neurotransmitter responsible for regulating most behaviors in the brain. An inefficient 5-HT synaptic function is often linked to various mental disorders. Primarily, membrane proteins controlling the expression and activity of 5-HT synthesis, storage, release, receptor activation, and inactivation are critical to 5-HT signaling in synaptic and extra-synaptic sites. Moreover, these signals represent information transmission across membranes. Although the lipid membrane environment is often viewed as fairly stable, emerging research suggests significant functional lipid−protein interactions with many synaptic 5-HT proteins. These protein−lipid interactions extend to almost all the primary lipid classes that form the plasma membrane. Collectively, these lipid classes and lipid−protein interactions affect 5-HT synaptic efficacy at the synapse. The highly dynamic lipid composition of synaptic membranes suggests that these lipids and their interactions with proteins may contribute to the plasticity of the 5-HT synapse. Therefore, this broader protein−lipid model of the 5-HT synapse necessitates a reconsideration of 5-HT's role in various associated mental disorders.

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Ceramide-mediation of diffusion in supported lipid bilayers

Cited by 2 publications

References 98 publications

Dissecting antibiotic effects on the cell envelope using bacterial cytological profiling: a phenotypic analysis starter kit

Dissecting antibiotic effects on the cell envelope using bacterial cytological profiling: a phenotypic analysis starter kit

Serotonin Signaling through Lipid Membranes

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