PERMEABILIZING ACTION OF FILIPIN-III ON MODEL MEMBRANES THROUGH A FILIPIN-PHOSPHOLIPID BINDING

被引:28
作者
MILHAUD, J
机构
[1] Laboratoire de Physique et Chimie Biomoléculaire (U.A. CNRS 198), Université Pierre et Marie Curie, Paris
关键词
POLYENE ANTIBIOTIC; FILIPIN; DIMYRISTOYLPHOSPHATIDYLCHOLINE; FLUORESCENCE PROBE; CARBOXYFLUORESCEIN; CALCEIN; PERMEABILIZATION; MODEL MEMBRANE; LIGHT SCATTERING; CD;
D O I
10.1016/0005-2736(92)90209-5
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The binding of the pentaene antibiotic filipin to egg-yolk phosphatidylcholine (EPC) and dimyristoylphosphatidylcholine (DMPC) unilamellar vesicles, has been studied by ultraviolet (UV) absorption and circular dichroism (CD). A stoichiometry of one molecule of filipin for five molecules of phospholipid was demonstrated by CD when phospholipids were in fluid phase. The similarity of the CD spectra with EPC and DMPC established a similar filipin-phospholipid assemblage in both membranes. We therefore postulated that filipin incorporation leads to the formation of gel-like domains in fluid EPC membranes as previously demonstrated for fluid DMPC membranes (Milhaud, J., Mazerski J., Bolard, J. and Dufourc, E.J. (1989) Eur. Biophys. J. 17, 151-158). The release of fluorescent probes (carboxyfluorescein (CF) or calcein (CC), entrapped in EPC small unilamellar vesicles (SUV), due to the action filipin, was followed by fluorescence and CD measurements concomitantly. The following observations were made. (1) The percentage of released probe, as a function of the filipin/phospholipid molar ratios, was the same whether or not membranes contained cholesterol. (2) The permeabilization of vesicles proceeded concomitantly with filipin-phospholipid binding while filipin-cholesterol binding leveled off. (3) The release of the content of vesicles occurred by an all-or-none mechanism leaving the depleted vesicles intact. From these observations and from the previous structural findings, a new interpretation of the action of filipin is proposed. Precluding any disruptive effect, inducement of permeability would result from the high intrinsic permeability of the interfacial region at the boundaries of the gel-like domains corresponding to the filipin-phospholipid aggregates. Additionally, we obtained the permeability coefficients for the anionic forms of CC and CF across EPC SUV, 0.6 . 10(-10)cm s-1 and 2 . 10(-10)cm s-1, respectively as compared to 2.5 . 10(-14)cm s-1 for the counterion Na+ (Hauser, H, Oldani, D. and Phillips, M.C. (1973) Biochemistry 12, 4507-4517).
引用
收藏
页码:307 / 318
页数:12
相关论文
共 61 条
[11]  
CANTOR CR, 1980, BIOPHYSICAL CHEM, V3, P859
[12]   KINETICS FOR DEVELOPMENT OF GRAMICIDIN-INDUCED ION PERMEABILITY IN UNILAMELLAR PHOSPHOLIPID-VESICLES [J].
CLEMENT, NR ;
GOULD, JM .
BIOCHEMISTRY, 1981, 20 (06) :1544-1548
[13]   PASSIVE ION PERMEABILITY OF LIPID-MEMBRANES MODELED VIA LIPID-DOMAIN INTERFACIAL AREA [J].
CRUZEIROHANSSON, L ;
MOURITSEN, OG .
BIOCHIMICA ET BIOPHYSICA ACTA, 1988, 944 (01) :63-72
[14]   PERMEABILITY OF LIPID BILAYERS TO WATER AND IONIC SOLUTES [J].
DEAMER, DW ;
BRAMHALL, J .
CHEMISTRY AND PHYSICS OF LIPIDS, 1986, 40 (2-4) :167-188
[15]   POLYENE ANTIBIOTIC-STEROL INTERACTIONS IN MEMBRANES OF ACHOLEPLASMA-LAIDLAWII CELLS AND LECITHIN LIPOSOMES .1. SPECIFICITY OF MEMBRANE-PERMEABILITY CHANGES INDUCED BY POLYENE ANTIBIOTICS [J].
DEKRUIJFF, B ;
GERRITSEN, WJ ;
OERLEMANS, A ;
DEMEL, RA ;
VANDEENE.LL .
BIOCHIMICA ET BIOPHYSICA ACTA, 1974, 339 (01) :30-43
[16]   POLYENE ANTIBIOTIC-STEROL INTERACTIONS IN MEMBRANES OF ACHOLEPLASMA-LAIDLAWII CELLS AND LECITHIN LIPOSOMES .3. MOLECULAR-STRUCTURE OF POLYENE ANTIBIOTIC-CHOLESTEROL COMPLEXES [J].
DEKRUIJFF, B ;
DEMEL, RA .
BIOCHIMICA ET BIOPHYSICA ACTA, 1974, 339 (01) :57-70
[17]   H-2 NMR EVIDENCE FOR ANTIBIOTIC-INDUCED CHOLESTEROL IMMOBILIZATION IN BIOLOGICAL MODEL MEMBRANES [J].
DUFOURC, EJ ;
SMITH, ICP .
BIOCHEMISTRY, 1985, 24 (10) :2420-2424
[18]  
DUFOURC EJ, 1986, SURFACTANTS SOLUTION, P897
[19]  
DUFOURC EJ, 1988, MEMBRANE BIOGENESIS, P177