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PMID: 8968598 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Effect of cholesterol and charge on pore formation in bilayer vesicles by a pH-sensitive peptide.

Biophysical journal ·Vol. 71 ·No. 6 ·1996-12-00 ·Pages 3288-301

Nicol F, Nir S, Szoka FC

Abstract

The effect of cholesterol on the bilayer partitioning of the peptide GALA (WEAALAEALAEALAEHLAEALAEALEALAA) and its assembly into a pore in large unilamellar vesicles composed of neutral and negatively charged phospholipids has been determined. GALA undergoes a conformational change from a random coil to an amphipathic alpha-helix when the pH is reduced from 7.0 to 5.0, inducing at low pH leakage of contents from vesicles. Leakage from neutral or negatively charged vesicles at pH 5.0 was similar and could be adequately explained by the mathematical model (Parente, R. A., S. Nir, and F. C. Szoka, Jr., 1990. Mechanism of leakage of phospholipid vesicle contents induced by the peptide GALA. Biochemistry. 29:8720-8728) which assumed that GALA becomes incorporated into the vesicle bilayer and irreversibly aggregates to form a pore consisting of 10 +/- 2 peptides. Increasing cholesterol content in the membranes resulted in a reduced efficiency of the peptide to induce leakage. Part of the cholesterol effect was due to reduced binding of the peptide to cholesterol-containing membranes. An additional effect of cholesterol was to increase reversibility of surface aggregation of the peptide in the membrane. Results could be explained and predicted with a model that retains the same pore size, i.e., 10 +/- 2 peptides, but includes reversible aggregation of the monomers to form the pore. Resonance energy transfer experiments using fluorescently labeled peptides confirmed that the degree of reversibility of surface aggregation of GALA was significantly larger in cholesterol-containing liposomes, thus reducing the efficiency of pore formation.

MeSH Terms
Amino Acid Sequence Cholesterol Dextrans Hydrogen-Ion Concentration Kinetics Lipid Bilayers/chemistry Models, Biological Models, Theoretical Molecular Sequence Data Peptides/chemical synthesis,chemistry Phosphatidylcholines/chemistry Phosphatidylglycerols/chemistry Protein Conformation Protein Structure, Secondary
Chemicals
Dextrans Lipid Bilayers Peptides Phosphatidylcholines Phosphatidylglycerols GALA peptide 1-palmitoyl-2-oleoylglycero-3-phosphoglycerol Cholesterol 1-palmitoyl-2-oleoylphosphatidylcholine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Nicol F
University of California, School of Pharmacy, Department of Biopharmaceutics, San Francisco 94143-0446, USA.
Nir S
Szoka F C
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Article Info
Journal
Biophysical journal
Abbr.
Biophys J
ISSN
0006-3495
DOI
10.1016/S0006-3495(96)79521-8
Published
1996-12-00
Pages
3288-301
Language
English
Region
United States
NLM ID
0370626
PMCID
PMC1233816
Subset
IM
Grants
NIDDK NIH HHS · DK46052 · United States
NIGMS NIH HHS · GM30163 · United States
External Links
PubMed source DOI: 10.1016/S0006-3495(96)79521-8 PMC Full Text View journal details
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