scholarly journals Fusion of phospholipid vesicles with planar phospholipid bilayer membranes. I. Discharge of vesicular contents across the planar membrane.

1980 ◽  
Vol 75 (3) ◽  
pp. 241-250 ◽  
Author(s):  
J Zimmerberg ◽  
F S Cohen ◽  
A Finkelstein
Keyword(s):  
Bilayer Membrane ◽  
Convincing Evidence ◽  
Water Soluble ◽  
Phospholipid Bilayer ◽  
Phospholipid Vesicles ◽  
Biological Phenomenon ◽  
Vesicle Membrane ◽  
Bilayer Membranes ◽  
Phospholipid Bilayer Membrane ◽  
Planar Membrane

Multilamellar phospholipid vesicles are introduced into the cis compartment on one side of a planar phospholipid bilayer membrane. The vesicles contain a water-soluble fluorescent dye trapped in the aqueous phases between the lamellae. If a vesicle containing n lamellae fuses with a planar membrane, an n-1 lamellar vesicle should be discharged into the opposite trans compartment, where it would appear as a discernible fluorescent particle. Thus, fusion events can be assayed by counting the number of fluorescent particles appearing in the trans compartment. In the absence of divalent cation, fusion does not occur, even after vesicles have been in the cis compartment for 40 min. When CaCl2 is introduced into the cis compartment to a concentration of greater than or equal to 20 mM, fusion occurs within the next 20 min; it generally ceases thereafter because of vesicle aggregation in the cis compartment. With approximately 3 x 10(8) vesicles/cm3 in the cis compartment, about 25-50 fusion events occur following CaCl2 addition. The discharge of vesicular contents across the planar membrane is the most convincing evidence of vesicle-membrane fusion and serves as a model for that ubiquitous biological phenomenon--exocytosis.

scholarly journals Fusion of phospholipid vesicles with planar phospholipid bilayer membranes. II. Incorporation of a vesicular membrane marker into the planar membrane.

1980 ◽  
Vol 75 (3) ◽  
pp. 251-270 ◽  
Author(s):  
F S Cohen ◽  
J Zimmerberg ◽  
A Finkelstein
Keyword(s):  
Divalent Cations ◽  
Phospholipid Bilayer ◽  
Phospholipid Vesicles ◽  
Osmotic Gradient ◽  
Bilayer Membranes ◽  
Fusion Procedure ◽  
Vesicular Membrane ◽  
Membrane Contact ◽  
Planar Membrane ◽  
Planar Membranes

Fusion of multilamellar phospholipid vesicles with planar phospholipid bilayer membranes was monitored by the rate of appearance in the planar membrane of an intrinsic membrane protein present in the vesicle membranes. An essential requirement for fusion is an osmotic gradient across the planar membrane, with the cis side (the side containing the vesicles) hyperosmotic to the opposite (trans) side; for substantial fusion rates, divalent cation must also be present on the cis side. Thus, the low fusion rates obtained with 100 mM excess glucose in the cis compartment are enhanced orders of magnitude by the addition of 5-10 mM CaCl2 to the cis compartment. Conversely, the rapid fusion rates induced by 40 mM CaCl2 in the cis compartment are completely suppressed when the osmotic gradient (created by the 40 mM CaCl2) is abolished by addition of an equivalent amount of either CaCl2, NaCl, urea, or glucose to the trans compartment. We propose that fusion occurs by the osmotic swelling of vesicles in contact with the planar membrane, with subsequent rupture of the vesicular and planar membranes in the region of contact. Divalent cations catalyze this process by increasing the frequency and duration of vesicle-planar membrane contact. We argue that essentially this same osmotic mechanism drives biological fusion processes, such as exocytosis. Our fusion procedure provides a general method for incorporating and reconstituting transport proteins into planar phospholipid bilayer membranes.

Design of artificial membrane transporters from gold nanoparticles with controllable hydrophobicity

2016 ◽  
Vol 191 ◽  
pp. 495-510 ◽  
Author(s):  
Marcin P. Grzelczak ◽  
Alexander P. Hill ◽  
Domagoj Belic ◽  
Dan F. Bradley ◽  
Casper Kunstmann-Olsen ◽  
...  
Keyword(s):  
Gold Nanoparticles ◽  
Phase Transfer ◽  
Aqueous Dispersion ◽  
Bilayer Membrane ◽  
Phospholipid Bilayer ◽  
Potassium Ions ◽  
Vesicle Membrane ◽  
Ligand Shell ◽  
Phospholipid Bilayer Membrane ◽  
Liquid Systems

Gold nanoparticles with variable hydrophobicity have been prepared in three different size regimes following established methods. The control of hydrophobicity was achieved by complexation of the 18-crown-6-CH2-thiolate ligand shell with potassium ions. Potassium dependent phase transfer of these particles from dispersion in water to chloroform was demonstrated, and the equilibrium partitioning of the particles in water–chloroform liquid/liquid systems was quantified by optical spectroscopy. The gradual complexation of the ligand shell with potassium ions was further monitored by zeta potential measurements. Potassium dependent insertion of nanoparticles into the phospholipid bilayer membrane of vesicles in aqueous dispersion has been demonstrated by cryogenic transmission electron microscopy (cryo-TEM). Nanoparticle-dependent potassium ion transport across the vesicle membrane has been established by monitoring the membrane potential with fluorescence spectroscopy using a potential sensitive dye.

scholarly journals Parameters affecting the fusion of unilamellar phospholipid vesicles with planar bilayer membranes.

1984 ◽  
Vol 98 (3) ◽  
pp. 1054-1062 ◽  
Author(s):  
F S Cohen ◽  
M H Akabas ◽  
J Zimmerberg ◽  
A Finkelstein
Keyword(s):  
Divalent Cation ◽  
Phospholipid Bilayer ◽  
Fusion Process ◽  
Phospholipid Vesicles ◽  
Osmotic Gradient ◽  
Planar Bilayer ◽  
Unilamellar Vesicles ◽  
Bilayer Membranes ◽  
Planar Membrane ◽  
Planar Membranes

It was previously shown (Cohen, F. S., J. Zimmerberg, and A. Finkelstein, 1980, J. Gen. Physiol., 75:251-270) that multilamellar phospholipid vesicles can fuse with decane-containing phospholipid bilayer membranes. An essential requirement for fusion was an osmotic gradient across the planar membrane, with the vesicle-containing (cis) side hyperosmotic with respect to the opposite (trans) side. We now report that unilamellar vesicles will fuse with "hydrocarbon-free" membranes subject to these same osmotic conditions. Thus the same conditions that apply to fusion of multilamellar vesicles with planar bilayer membranes also apply to fusion of unilamellar vesicles with these membranes, and hydrocarbon is not required for the fusion process. If the vesicles and/or planar membrane contain negatively charged lipids, divalent cation (approximately 15 mM Ca++) is required in the cis compartment (in addition to the osmotic gradient across the membrane) to obtain substantial fusion rates. On the other hand, vesicles made from uncharged lipids readily fuse with planar phosphatidylethanolamine planar membranes in the near absence of divalent cation with just an osmotic gradient. Vesicles fuse much more readily with phosphatidylethanolamine-containing than with phosphatidylcholine-containing planar membranes. Although hydrocarbon (decane) is not required in the planar membrane for fusion, it does affect the rate of fusion and causes the fusion process to be dependent on stirring in the cis compartment.

scholarly journals Evaluation of the correlation between porphyrin accumulation in cancer cells and functional positions for application as a drug carrier

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Koshi Nishida ◽  
Toshifumi Tojo ◽  
Takeshi Kondo ◽  
Makoto Yuasa
Keyword(s):  
Cancer Cells ◽  
Drug Carrier ◽  
Bilayer Membrane ◽  
Phospholipid Bilayer ◽  
Membrane Permeation ◽  
Porphyrin Derivatives ◽  
Phospholipid Bilayer Membrane ◽  
Meso Position ◽  
Tumor Accumulation ◽  
Binding Position

AbstractPorphyrin derivatives accumulate selectively in cancer cells and are can be used as carriers of drugs. Until now, the substituents that bind to porphyrins (mainly at the meso-position) have been actively investigated, but the effect of the functional porphyrin positions (β-, meso-position) on tumor accumulation has not been investigated. Therefore, we investigated the correlation between the functional position of substituents and the accumulation of porphyrins in cancer cells using cancer cells. We found that the meso-derivative showed higher accumulation in cancer cells than the β-derivative, and porphyrins with less bulky substituent actively accumulate in cancer cells. When evaluating the intracellular distribution of porphyrin, we found that porphyrin was internalized by endocytosis and direct membrane permeation. As factors involved in these two permeation mechanisms, we evaluated the affinity between porphyrin-protein (endocytosis) and the permeability to the phospholipid bilayer membrane (direct membrane permeation). We found that the binding position of porphyrin affects the factors involved in the transmembrane permeation mechanisms and impacts the accumulation in cancer cells.

Synthetic, Sodium-Ion-Conducting Tris(Macrocycle) Channels that Function in a Phospholipid Bilayer Membrane: An Overview

2000 ◽  
Vol 12 (1) ◽  
pp. 13-22
Author(s):  
Stephen L. De Wall ◽  
Eric S. Meadows ◽  
Clare L. Murray ◽  
Hossein Shabany ◽  
George W. Gokel
Keyword(s):  
Bilayer Membrane ◽  
Sodium Ion ◽  
Phospholipid Bilayer ◽  
Phospholipid Bilayer Membrane ◽  
Ion Conducting

scholarly journals Fusion of phospholipid vesicles with a planar membrane depends on the membrane permeability of the solute used to create the osmotic pressure.

1989 ◽  
Vol 93 (2) ◽  
pp. 201-210 ◽  
Author(s):  
F S Cohen ◽  
W D Niles ◽  
M H Akabas
Keyword(s):  
Osmotic Pressure ◽  
Lipid Bilayer ◽  
Lipid Membrane ◽  
Contact Region ◽  
Companion Paper ◽  
Phospholipid Vesicles ◽  
Osmotic Gradient ◽  
Vesicle Membrane ◽  
Permeable Channel ◽  
Planar Membrane

Phospholipid vesicles fuse with a planar membrane when they are osmotically swollen. Channels in the vesicle membrane are required for swelling to occur when the vesicle-containing compartment is made hyperosmotic by adding a solute (termed an osmoticant). We have studied fusion using two different channels, porin, a highly permeable channel, and nystatin, a much less permeable channel. We report that an osmoticant's ability to support fusion (defined as the magnitude of osmotic gradient necessary to obtain sustained fusion) depends on both its permeability through lipid bilayer as well as its permeability through the channel by which it enters the vesicle interior. With porin as the channel, formamide requires an osmotic gradient about ten times that required with urea, which is approximately 1/40th as permeant as formamide through bare lipid membrane. When nystatin is the channel, however, fusion rates sustained by osmotic gradients of formamide are within a factor of two of those obtained with urea. Vesicles containing a porin-impermeant solute can be induced to swell and fuse with a planar membrane when the impermeant bathing the vesicles is replaced by an isosmotic quantity of a porin-permeant solute. With this method of swelling, formamide is as effective as urea in obtaining fusion. In addition, we report that binding of vesicles to the planar membrane does not make the contact region more permeable to the osmoticant than is bare lipid bilayer. In the companion paper, we quantitatively account for the observation that the ability of a solute to promote fusion depends on its permeability properties and the method of swelling. We show that the intravesicular pressure developed drives fusion.

scholarly journals Diffusion in phospholipid bilayer membranes: dual-leaflet dynamics and the roles of tracer–leaflet and inter-leaflet coupling

2014 ◽  
Vol 470 (2167) ◽  
pp. 20130843 ◽  
Author(s):  
Reghan J. Hill ◽  
Chih-Ying Wang
Keyword(s):  
Lateral Diffusion ◽  
Bilayer Membrane ◽  
Tracer Diffusion ◽  
Phospholipid Bilayer ◽  
Supported Membranes ◽  
Bilayer Membranes ◽  
Lubrication Film ◽  
Lipid Assemblies ◽  
Slip Coefficients ◽  
Made In

A variety of observations—sometimes controversial—have been made in recent decades when attempting to elucidate the roles of interfacial slip on tracer diffusion in phospholipid membranes. Evans–Sackmann theory (1988) has furnished membrane viscosities and lubrication-film thicknesses for supported membranes from experimentally measured lateral diffusion coefficients. Similar to the Saffman and Delbrück model, which is the well-known counterpart for freely supported membranes, the bilayer is modelled as a single two-dimensional fluid. However, the Evans–Sackman model cannot interpret the mobilities of monotopic tracers, such as individual lipids or rigidly bound lipid assemblies; neither does it account for tracer–leaflet and inter-leaflet slip. To address these limitations, we solve the model of Wang and Hill, in which two leaflets of a bilayer membrane, a circular tracer and supports are coupled by interfacial friction, using phenomenological friction/slip coefficients. This furnishes an exact solution that can be readily adopted to interpret the mobilities of a variety of mosaic elements—including lipids, integral monotopic and polytopic proteins, and lipid rafts—in supported bilayer membranes.

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