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WIND-PVPA: Water/Ion NMR Detected PVPA to assess lipid barrier integrity in vitro through quantification of passive water- and ion transport

Permanent lenke
https://hdl.handle.net/10037/25229
DOI
https://doi.org/10.1016/j.bbamem.2022.183911
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article.pdf (5.455Mb)
Publisert versjon (PDF)
Dato
2022-03-22
Type
Journal article
Tidsskriftartikkel
Peer reviewed

Forfatter
Rainsford, Philip Ben; Sarre, Birta R.; Falavigna, Margherita; Brandsdal, Bjørn Olav; Flaten, Gøril Eide; Jakubec, Martin; Isaksson, Johan Mattias
Sammendrag
Water/Ion NMR Detected – Phospholipid Vesicle Permeability Assay (WIND-PVPA), is presented as a novel, straightforward and automatable method to assess lipid barrier integrity in vitro. The apparent permeability constants of water- and ions across the PVPA barriers are determined in a one-pot experiment under the influence of membrane-active guest molecules. NMR spectroscopy is used to quantify the water directly (D2O) and the ions indirectly (complexed with EDTA) as a function of time. WIND-PVPA is demonstrated using four anti-microbial peptides, to show that membrane active molecules can be differentiated by their disruptive influence on the PVPA system. The results obtained are compared with explicit molecular dynamics simulations of lipid bilayers, AMPs, water and salt, where the motions of all individual water molecules relative to the lipid bilayer are monitored over the course of the simulations, allowing the calculation of theoretical apparent permeability constants of the corresponding single bilayer systems. Proof-of-principle is presented that WIND-PVPA can be used to evaluate the lipid barrier destabilizing effect of active guest molecules by measuring changes in passive water- and ion permeabilities upon exposure. The method is highly flexible in terms of barrier composition, choice of probes and membrane active compounds.
Er en del av
Rainsford, P. (2022). Expanding the toolbox for the study of antimicrobial peptides. (Doctoral thesis). https://hdl.handle.net/10037/25236
Forlag
Elsevier
Sitering
Rainsford, Sarre, Falavigna, Brandsdal, Flaten, Jakubec, Isaksson. WIND-PVPA: Water/Ion NMR Detected PVPA to assess lipid barrier integrity in vitro through quantification of passive water- and ion transport. Biochimica et Biophysica Acta - Biomembranes. 2022;1864(7)
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