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PMID: 10653794 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Effect of phospholipid composition on an amphipathic peptide-mediated pore formation in bilayer vesicles.

Biophysical journal ·Vol. 78 ·No. 2 ·2000-02-00 ·Pages 818-29

Nicol F, Nir S, Szoka FC

Abstract

To better understand the influence of phospholipid acyl-chain composition on the formation of pores by cytotoxic amphipathic helices in biological membranes, the leakage of aqueous contents induced by the synthetic peptide GALA (WEAALAEALAE ALAEHLAEALAEALEALAA) from large unilamellar phospholipid vesicles of various compositions has been studied. Peptide-mediated leakage was examined at pH 5.0 from vesicles made of phosphatidylcholine (PC) and phosphatidylglycerol (PG) with the following acyl-chain compositions: 1-palmitoyl-2-oleoyl (PO), 1,2-dioleoyl (DO), 1, 2-dielaidoyl (DE), and 1,2-dipetroselinoyl (DPe). A mathematical model predicts and simulates the final extents of GALA-mediated leakage of 1-aminonaphthalene-3,6,8-trisulfonic acid (ANTS) and p-xylene-bis-pyridinium bromide (DPX) from 1-palmitoyl-2-oleoyl-phosphatidylcholine/1-palmitoyl-2-oleoyl-phospha tidylglycerol (POPC/POPG) and 1, 2-dielaidoyl-sn-glycero-3-phosphocholine/1, 2-dielaidoyl-phosphatidylglycerol (DEPC/DEPG) liposomes at pH 5.0 as a function of peptide concentration in the bilayer, by considering that GALA pores responsible for this leakage have a minimum size of 10 +/- 2 monomers and are formed by quasiirreversible aggregation of the peptide. With the phospholipid acyl-chain compositions tested, GALA-induced ANTS/DPX leakage follows the rank order POPC/POPG approximately DEPC/DEPG > DPePC/DPePG > DOPC/DOPG. Results from binding experiments reveal that this reduced leakage from DOPC/DOPG vesicles cannot be explained by a reduced binding affinity of the peptide to these membranes. As shown by monitoring the leakage of a fluorescent dextran, an increase in the minimum pore size also does not explain the reduction in ANTS/DPX leakage. The data suggest that surface-associated GALA monomers or aggregates are stabilized in bilayers composed of phospholipids containing a cis unsaturation per acyl chain (DO and DPe), while transbilayer peptide insertion is reduced. GALA-induced ANTS/DPX leakage is also decreased when the vesicles contain phosphatidylethanolamine (PE). This lends further support to the suggestion that factors stabilizing the surface state of the peptide reduce its insertion and subsequent pore formation in the bilayer.

MeSH Terms
4-Chloro-7-nitrobenzofurazan Amino Acid Sequence Animals Dextrans Fluorescent Dyes Ion Channels/chemistry Kinetics Liposomes/chemistry Molecular Sequence Data Naphthalenes Peptides/chemistry,pharmacology Permeability Phospholipids/chemistry Pyridinium Compounds Xanthenes
Chemicals
Dextrans Fluorescent Dyes Ion Channels Liposomes Naphthalenes Peptides Phospholipids Pyridinium Compounds Xanthenes GALA peptide N,N'-4-xylylenebis(pyridinium) Texas red 8-amino-1,3,6-naphthalenetrisulfonic acid 4-Chloro-7-nitrobenzofurazan
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Nicol F
School of Pharmacy, University of California, San Francisco, California 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(00)76639-2
Published
2000-02-00
Pages
818-29
Language
English
Region
United States
NLM ID
0370626
PMCID
PMC1300684
Subset
IM
Grants
NIDDK NIH HHS · DK 46052 · United States
External Links
PubMed source DOI: 10.1016/S0006-3495(00)76639-2 PMC Full Text View journal details
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