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Effects of bilirubin molecular species on membrane dynamic properties of human erythrocyte membranes: a spin label electron paramagnetic resonance spectroscopy study

dc.contributor.authorBrito, Maria Alexandra
dc.contributor.authorBrondino, Carlos D.
dc.contributor.authorMoura, José J. G.
dc.contributor.authorBrites, Dora
dc.date.accessioned2017-08-07T12:15:14Z
dc.date.available2017-08-07T12:15:14Z
dc.date.issued2001
dc.description.abstractUnconjugated bilirubin is a neurotoxic pigment that interacts with membrane lipids. In this study we used electron paramagnetic resonance and the spin labels 5-, 7-, 12-, and 16-doxyl-stearic acid (DSA) to evaluate the depth of the hydrocarbon chain at which interaction of bilirubin preferentially occurs. In addition, we used different pH values to determine the molecular species involved. Resealed right-side-out ghosts were incubated (1–60 min) with bilirubin (3.4–42.8 μM) at pH 7.0, 7.4, and 8.0. Alterations of membrane dynamic properties were maximum after 15 min of incubation with 8.6 μM bilirubin at pH 7.4 and were accompanied by a significant release of phospholipids. Interestingly, concentrations of bilirubin up to 42.8 μM and longer incubations resulted in the elution of cholesterol and further increased that of phospholipids while inducing less structural alterations. Variation of the pH values from 8.0 to 7.4 and 7.0, under conditions of maximum perturbation, led to a change from an increased to a diminished polarity sensed by 5-DSA. Conversely, a progressive enhancement in fluidity was reported by 7-DSA, followed by 12- and 16-DSA. These results indicate that bilirubin while enhancing membrane lipid order at C-5 simultaneously has disordering effects at C-7. Furthermore, recovery of membrane dynamics after 15 min of bilirubin exposure along with the release of lipids is compatible with a membrane adaptive response to the insult. In addition, our data provide evidence that uncharged diacid is the species primarily interacting with the membrane as perturbation is favored by acidosis, a condition frequently associated with hyperbilirubinemia in premature and severely ill infants.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationArchives of Biochemistry and Biophysics. 2001;387(1):57-65pt_PT
dc.identifier.doihttps://doi.org/10.1006/abbi.2000.2210pt_PT
dc.identifier.issn0003-9861
dc.identifier.urihttp://hdl.handle.net/10451/28636
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.relation.publisherversionhttp://www.sciencedirect.com/science/article/pii/S0003986100922100pt_PT
dc.subjectbilirubinpt_PT
dc.subjectelectron paramagnetic resonancept_PT
dc.subjecthuman erythrocytespt_PT
dc.subjectlipidspt_PT
dc.subjectmembrane ftuiditypt_PT
dc.subjectmembrane orderpt_PT
dc.subjectspin labelspt_PT
dc.titleEffects of bilirubin molecular species on membrane dynamic properties of human erythrocyte membranes: a spin label electron paramagnetic resonance spectroscopy studypt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage65pt_PT
oaire.citation.issue1pt_PT
oaire.citation.startPage57pt_PT
oaire.citation.titleArchives of Biochemistry and Biophysicspt_PT
oaire.citation.volume387pt_PT
rcaap.rightsrestrictedAccesspt_PT
rcaap.typearticlept_PT

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