Early stages of oxidative stress-induced membrane permeabilization: A neutron reflectometry study

Hillary L. Smith, Michael C. Howland, Alan W. Szmodis, Li Qijuan, Luke L. Daemen, Atul N. Parikh, Jaroslaw Majewski

Research output: Contribution to journalArticlepeer-review

66 Scopus citations

Abstract

Neutron reflectometry was used to probe in situ the structure of supported lipid bilayers at the solid-liquid interface during the early stages of UV-induced oxidative degradation. Single-component supported lipid bilayers composed of gel phase, dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), and fluid phase, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), phospholipids were exposed to low-dose oxidative stress generated by UV light and their structures were examined by neutron reflectometry. An interrupted illumination mode, involving exposures in 15 min increments with 2 h intervals between subsequent exposures, and a continuous mode involving a single 60 (or 90) min exposure period were employed. In both cases, pronounced differences in the structure of the lipid bilayer after exposure were observed. Interrupted exposure led to a substantial decrease in membrane coverage but preserved its total thickness at reduced scattering length densities. These results indicate that the initial phase during UV-induced membrane degradation involves the formation of hydrophilic channels within the membrane. This is consistent with the loss of some lipid molecules we observe and attendant reorganization of residual lipids forming hemimicellar edges of the hydrophilic channels. In contrast, continuous illumination produced a graded interface of continuously varied scattering length density (and hence hydrocarbon density) extending 100-150 Å into the liquid phase. Exposure of a DPPC bilayer to UV light in the presence of a reservoir of unfused vesicles showed low net membrane disintegration during oxidative stress, presumably because of surface back-filling from the bulk reservoir. Chemical evidence for membrane degradation was obtained by mass spectrometry and Fourier transform infrared spectroscopy. Further evidence for the formation of hydrophilic channels was furnished by fluorescence microscopy and imaging ellipsometry data.

Original languageEnglish
Pages (from-to)3631-3638
Number of pages8
JournalJournal of the American Chemical Society
Volume131
Issue number10
DOIs
StatePublished - Mar 18 2009
Externally publishedYes

Funding

FundersFunder number
National Eye InstitutePN2EY016570
National Institute of General Medical SciencesT32GM008799

    Fingerprint

    Dive into the research topics of 'Early stages of oxidative stress-induced membrane permeabilization: A neutron reflectometry study'. Together they form a unique fingerprint.

    Cite this