Lateral diffusion and phase separation in two-dimensional solutions of polymerized butadiene lipid in dimyristoylphosphatidylcholine bilayers. A photobleaching and freeze fracture study

Gaub, H.; Sackmann, E.; Büschl, R.; Ringsdorf, Helmut

doi:10.1016/s0006-3495(84)84215-0

Cited by 71 publications

(50 citation statements)

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“…The existence of lateral and/or transverse lipid domains in membranes is often inferred from the properties of fluorescent lipids incorporated into model systems (1)(2)(3) or in cell membranes (4)(5)(6)(7)(8). Domains are generally associated with regions of distinct fluidity, sometimes corresponding to a pure gel or a pure liquid-crystalline state.…”

mentioning

confidence: 99%

Phospholipid transmembrane domains and lateral diffusion in fibroblasts.

Chahine

Cribier

Devaux

1993

Proc. Natl. Acad. Sci. U.S.A.

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The lateral diffusion of fluorescent phospholipids in cultured Chinese hamster lung fibroblasts was examined by modulated fringe pattern photobleaching. When cells were labeled and maintained at 7C, the fluorescence remained localized at the plasma membrane. N-[6-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl-amino)caproyllsphingosylphosphocholine (C6-NBD-SphPCho) and 1-acyl-2-[6-(7-nitrobenz-2-oxa-1,3-diazol-4-yl-amino)caproylJ phosphatidylcholine (C6-NBDPtdCho) both diffused with the same apparent lateral diffusion coefficient (D, 0.3 x 10-9 cm2/s). By contrast, the phosphatidylserine derivative {1-acyl-2-[6-(7-nitrobenz-2-oxa-1,3-diazol-4-yl-amino)caproylJ phosphatidylserine (C6-NBD-PtdSer)} gave rise to two diffusional components: a slow component, D1, analogous to that measured with the cholinecontaining lipids, and a fast component (D2 2 x 10-9 cm2/s).The fast component only exists in ATP-containing cells. It was shown to be associated with C6-NBD-PtdSer translocated to the inner leaflet. This indicates that the two leaflets form very different membranous domains. At higher temperature, the same difference in mobility was observed between the cholinecontaining lipids and the aminolipid. However, with C6-NBDSphPCho, a fraction of very slowly diffusing or quasiimmobilized probes gradually appeared with time. This could be attributed to sphingomyelin located in small organelles after internalization. From the amplitude of this component registered at different intervals, we calculated that =50% of the plasma membrane sphingomyelin is recycled in less than 30 min in Chinese hamster fibroblasts by an ATP-and microtubule-dependent process.The existence of lateral and/or transverse lipid domains in membranes is often inferred from the properties of fluorescent lipids incorporated into model systems (1-3) or in cell membranes (4-8). Domains are generally associated with regions of distinct fluidity, sometimes corresponding to a pure gel or a pure liquid-crystalline state. However, domains do not automatically imply lipid phase immiscibility per se but may also be delimited by barriers. For example, tight junctions restrict the lateral diffusion of lipids in polarized cells (9), whereas protein clusters would restrict that of plastoquinones in chloroplasts (10). Similarly, the hydrophobic core of all membranes constitutes a barrier to the passage of phospholipids from one leaflet to the other. In red blood cells, ESR and fluorescence techniques were used to demonstrate a fluidity difference between inner and outer leaflets of the plasma membrane without implying gel formation (11-13). The difference was attributed to the specific lipid composition of each leaflet, in particular to the greater degree ofunsaturation ofthe lipid chains in the inner monolayer (14).All eukaryotic cells seem to have a plasma membrane with an asymmetrical phospholipid distribution (for a review, see ref. 15). In the present article, we show that the asymmetrical transverse lipid composition of V79 cultured Chinese hamster fibroblast membranes...

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confidence: 99%

Phospholipid transmembrane domains and lateral diffusion in fibroblasts.

Chahine

Cribier

Devaux

1993

Proc. Natl. Acad. Sci. U.S.A.

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“…21, No. 8,1989 Reconstitution of Liposomes frofJl DODPC Polymers 18 Small unilamellar liposomes prepared by sonication method were polymerized with AIBN at 60°C for 12 h. Diene groups in 1-acyl chains were selectively polymerized by this procedure. 11 The linear (comb like) DODPC polymers (M" = 1.67 x 10 4 by GPC calculated from polystyrene standard sample 19 ) were obtained by dissolving the liposomes lyophilized from water into benzene.…”

Section: Preparation Of Poly»jerized Liposomementioning

confidence: 99%

“…4 CF release measurement, electron microscopy and light-scattering measurement were often applied to analyze the stability of polymerized liposomes. 5 -7 There were a few methods to evaluate phase transition behavior of polymerized liposome membrane such as photobleaching 8 and DSC measurement. 9 * To whom all correspondence should be addressed.…”

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Study on the Phase Transition Behavior of Polymerized Liposomes through the Interaction of Diene-Groups in Their Acyl Chains

Takeoka

Sakai

Ohno

et al. 1989

Polym J

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ABSTRACT:I ,2-Bis(2,4-octadecadienoyl)-sn-glycero-3-phosphorylcholine (DODPC), which had a diene group in 2,4-position of I-and 2-acyl chains, was used to detect phase transition from shift of absorption maximum Uma,l due to the change of interaction of diene groups. Selective polymerization of 1-acyl chains of DODPC with AIBN gave linear phospholipid polymers which had unreacted diene groups in 2-acyl chains. From the analysis of ).m., of thus selectivelypolymerized liposomes, the phase transition behavior was detected clearly. We also studied the phase transition behavior of liposomes of which either layer of bilayer was selectively polymerized. Reconstitution of polymerized DODPC liposomes could also be evaluated from the temperature dependence of ).m.,· The molecular packing of the polymeric liposomes was disturbed by reconstitution because of lower freedom of phospholipid polymers on reconstitution process. KEY WORDSLiposome / Phase Transition / Polymerized Liposomes / Reconstitution / Diene-Containing Lipids / Absorption Maximum / The liposome has hitherto been studied for the application of drug delivery system because both hydrophobic bilayer and an inner aqueous phase can be used for encapsulation of functional molecules. 1 · 2 Liposomes, which can release entrapped materials in response to the onter stimuli, were designed and studied in physiological conditions. 2 Liposomes tend to aggregate and fuse after storage at low temperature. 3 They are not stable enough and sometimes release encapsulated molecules easily in vivo or in vitro. 2 Polymerization of liposomes components is a potent technique to prepare the stable liposomes. We succeeded in preparing polymerized mixed liposomes which released encapsulated molecules in response to physical or chemical stimuli. 4 CF release measurement, electron microscopy and light-scattering measurement were often applied to analyze the stability of polymerized liposomes. 5 -7 There were a few methods to evaluate phase transition behavior of polymerized liposome membrane such as photobleaching 8 and DSC measurement. 9* To whom all correspondence should be addressed.We have already used amphiphilic porphynato zinc, 3,8,13, 17-tetramethyl-7, 12-dicarboxy-2, 18-bis( octadecylpropinate )porphynato zinc (DCPZn) as a membrane probe for the detection of its phase transition temperature. 10 The fluorescence intensity changed in relation to the change of dissociation-association equilibrium of the amphiphilic porphyrins at temperatures below and above the phase transition temperature. It was however impossible to detect the phase transition of polymerized liposomes with this method, because these

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“…With such lipids it may be possible to create special membrane domains, which could conceivably supply information about protein-lipid and lipid-lipid interactions. Freeze-fracture studies with such lipids have begun, but only in model systems (Gaub et al, 1984;Leaver et al, 1983). Another series of virtually unexploited methods is the application of controlled decoration with water or heavy metals (discussed in Sections I11 and lVj).…”

Section: Lipid Cytochemistrymentioning

confidence: 99%

A survey of ultra‐rapid cryofixation methods with particular emphasis on applications to freeze‐fracturing, freeze‐etching, and freeze‐substitution

Menco

1986

J. Elec. Microsc. Tech.

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Lateral diffusion and phase separation in two-dimensional solutions of polymerized butadiene lipid in dimyristoylphosphatidylcholine bilayers. A photobleaching and freeze fracture study

Cited by 71 publications

References 15 publications

Phospholipid transmembrane domains and lateral diffusion in fibroblasts.

Phospholipid transmembrane domains and lateral diffusion in fibroblasts.

Study on the Phase Transition Behavior of Polymerized Liposomes through the Interaction of Diene-Groups in Their Acyl Chains

A survey of ultra‐rapid cryofixation methods with particular emphasis on applications to freeze‐fracturing, freeze‐etching, and freeze‐substitution

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