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Biomedical subjects

W Knoll

Publications and source records attributed to W Knoll.

14 recordsLinked to original sources
PubMed1992

Long-range attraction and molecular rearrangements in receptor-ligand interactions.

A surface force apparatus was used to measure a long-range attractive protein-ligand force at separations D less than 85 angstroms. This force may effectively "steer" ligand trajectories, resulting in a greater than 27-fold enhancement of the association rate. A much stronger specific attraction is measured at contact (D less than 4 angstroms). A sevenfold increase in intermembrane adhesion resulted from increased lateral mobility of the receptors and molecular rearrangements in membranes above the solid-fluid transition temperature.

Bacterial Proteins↗
PubMed1992

Streptavidin binding observed with an atomic force microscope.

An atomic force microscope (AFM) was used to investigate a specific recognition reaction: the binding of streptavidin to a biotinylated lipid bilayer. Prior to the recognition reaction, the phase coexistence of the lipid bilayer was clearly observed: fluid domains were lower than the crystalline domains. After introducing to the bilayer a very dilute solution of streptavidin to give a final concentration of approximately 0.5 microM, the recognition reaction was imaged in real time. Several hours later, we observed a contrast reversal, i.e., the previously lower fluid domains grew so much in height that they became higher than the crystalline domains. We found that the streptavidin molecules bound almost exclusively to the biotin in the fluid domain (less than 0.25% coverage of the crystalline domains). The apparent structure of the few streptavidin molecules bound to the crystalline domain of the bilayer is shown to depend on the applied force. Finally, in a 2-dimensional quasi-crystal in which the streptavidin molecules were compressed at the air-water interface molecular resolution was achieved.

Bacterial Proteins↗
PubMed1992

Streptavidin binding to biotinylated lipid layers on solid supports. A neutron reflection and surface plasmon optical study.

Neutron reflection and surface plasmon optical experiments have been performed to evaluate structural data of the interfacial binding reaction between the protein streptavidin and a solid-supported lipid monolayer partly functionalized by biotin moieties. Since both experimental techniques operate in a total internal reflection geometry at a substrate/solution interface, identical sample architectures allow for a direct comparison between the results obtained with these two recently developed methods. It is found that a monomolecular layer of dipalmitoyllecithin doped with 5 mol% of a biotinylated-phosphatidylethanolamine shows a thickness of d1 approximately (3.4 +/- 0.5) nm. Binding of streptavidin to the biotin groups results in an overall layer thickness of d = (5.9 + 0.5) nm that demonstrates the formation of a well-ordered protein monolayer with the (biotin+spacer) units of the functionalized lipids being fully embedded into the binding pocket of the proteins. It is demonstrated by model calculations that a more detailed picture of the internal structure of this supramolecular assembly can only be obtained if one uses deuterated lipid molecules, thus generating a high contrast between individual layers.

Bacterial Proteins↗
PubMed1991

Measurement of ligand-receptor interactions.

One distinguishing feature of "life" is that the physical forces between biological molecules and membrane surfaces are often highly specific, in contrast to nonspecific interactions such as van der Waals, hydrophobic, and electrostatic (Coulombic) forces. We have used the surface-forces-apparatus technique to study the specific "lock and key" or "ligand-receptor" interaction between two model biomembrane surfaces in aqueous solution. The membranes were lipid bilayers supported on mica surfaces; one carrying streptavidin receptors, the other exposing biotin ligand groups. We found that, although no unusual or specific interaction occurs between two avidin or two biotin surfaces, an avidin and a biotin surface exhibit a very strong, very short-range (less than 1 nm) attraction and that the binding mechanism involves equally specific molecular rearrangements. The results also show that highly specific biological interactions such as are involved in immunological recognition and cell-cell contacts may be studied at the molecular level and in real time by the surface-forces-apparatus technique.

Aluminum Silicates↗
PubMed1991

Lateral order in binary lipid alloys and its coupling to membrane functions.

Densitometry, Raman spectroscopy and small angle neutron scattering are employed to elucidate the miscibility behavior of lipid mixtures organized as liposomal dispersions. First, temperature-composition-phase diagrams for several binary alloys of dialkyl-lecithins differing in chain lengths by an increasing number of CH2-groups are derived. A mixture of dimyristoyllecithin and distearoyllecithin (delta CH2 = 4) shows a peritectic phase behavior with a miscibility gap in the gel state. In the fluid phase, at high enough temperatures, homogeneous mixtures of the two components are formed at all molar ratios. However, upon approaching a critical point by either lowering the temperature or increasing the hydrostatic pressure, critical concentration fluctuations are observed. If one component of a binary mixture is charged, electrostatic interactions can be used to induce phase separation at constant temperature. This is demonstrated for Ca2(+)-driven demixing in alloys of lecithin and negatively charged phosphatidylglycerol. The influence of the various concepts for the induction of lateral structure formation in lipid membranes on integral functional units like ionophores is demonstrated by analysing the single channel current fluctuations of gramicidin in bimolecular lipid membranes. Ca2+, as well as polyelectrolyte-induced phase separations are shown and discussed as examples for the important (lateral) order-function relationship in biomembranes.

Electrochemistry↗
PubMed1990

pH-control of the miscibility properties of a binary lipid alloy and its influence on the ion transport by gramicidin.

We studied the coupling of a membrane function (the transport of ions by the pore forming polypeptide gramicidin) to chemically driven phase changes in black membranes of binary lipid mixtures. In particular, we investigated the influence of the aqueous pH value on the fluid-fluid demixing effect of Ca2+ to phosphatidylcholine/phosphatidylglycerol bilayers. It is found that one can switch, under certain conditions, between a homogeneously mixed and a phase separated membrane by changing the pH. We interpret this as being caused by the change in the degree of dissociation of one of the lipid components.

Electric Conductivity↗
PubMed1989

Phase separation in bimolecular mixed lipid membranes induced by polylysine.

We demonstrate, for the first time, polylysine-induced phase separation in a bimolecular lipid membrane of a lecithin/phosphatidylglycerol-mixture by analysing the single channel current fluctuations of gramicidin. The bimodal conductance histograms are direct evidence for the incorporation of the transport system into the two coexisting phases of different composition.

Dimyristoylphosphatidylcholine↗
PubMed1989

Lateral order in mixed lipid bilayers and its influence on ion translocation by gramicidin: a model for the structure-function relationship in membranes.

The temperature-composition phase diagram of dimyristoylphosphatidylcholine and dipentadecylphosphatidylglycerol (DiC15PG) was determined by mass densitometry. For a mixture containing 30 mol% DiC15PG, the homogeneous distribution of the 2 components is demonstrated in the fluid state at T = 35 degrees C by small-angle neutron scattering in combination with the inverse contrast variation method. By the same technique, the coexistence of fluid and condensed phases at T = 23.3 degrees C could be shown in agreement with the densitometric data. Furthermore, it is demonstrated that Ca++ induces, even at T = 35 degrees C, separation into 2 fluid phases. A corresponding phase separation is found in bimolecular lipid membranes ("black films") by analysis of the single-channel conductance fluctuations of gramicidin A incorporated into an equimolarly mixed membrane of neutral lecithin and charged phosphatidic acid. The results are discussed as primary examples on the model-membrane level for the important structure-function relationship of biomembranes.

Calcium↗
PubMed1986

Direct evidence for Ca++-induced lateral phase separation in black membranes of lipid mixtures by the analysis of gramicidin A single-channels.

Single-channel conductance fluctuations are analysed for gramicidin A incorporated into binary-mixed black lipid membranes of charged phosphatidic acid and neutral lecithin in different molar ratios. At very low Ca++ concentrations in the electrolyte (i.e. in the presence of EDTA) homogeneous lipid mixtures are identified through their conductance and life time probability distributions for integral gramicidin pores. As for the pure lipid components, the conductance histograms each show a single maximum with regular width and for all channels a single mean lifetime is found. For Ca++-levels (10(-6) - 10(-5) M) that are close to the critical demixing concentration (approximately 10(-4) M) unusually broad conductance distributions and reduced lifetimes are found provided the PC content, chi, of the membrane is close to the critical mixture (chi crit approximately equal to 0.5). We interpret this as a first example of the coupling of a membrane function (the transport of ions) to a lipid matrix with locally fluctuating composition close to a critical demixing point. For cCA++ = 10(-2) M the conductance histogram of gramicidin A in an equimolar mixture of PA and PC shows two well-separated maxima. A correlation analysis between conductance and lifetime of the single pores shows that the two channel populations also differ significantly in their mean channel lifetime, tau *. This finding is interpreted as being direct evidence for Ca++-induced lateral phase separation in black lipid membranes, as has been postulated recently.

Calcium↗
PubMed1977

Temperature-jump experiments on thin lipid membranes in the presence of valinomycin.

Temperature jump relaxation experiments on planar lipid membranes in the presence of valinomycin were performed using the absorption of a strong light flash as an energy source for the generation of the T-jump. The relaxation of the current carried by valinomycin/Rb+ complexes was measured. The results were interpreted on the basis of a transport model which was also analyzed by voltage jump relaxation experiments. The study shows that the application of the T-jump technique provides valuable information about transport kinetics as well as the dynamics of the membrane structure. At the given experimental conditions the relaxation of the current is believed to reflect a temperature-dependent transition of the membrane to a new conformational state of low order. The relaxation could be resolved with the present technique only at low temperatures and for membranes of high microviscosity.

Biological Transport↗
PubMed1976

Optical and electrical studies on dansyllysine-valinomycin in thin lipid membranes.

Dansyllysine-valinomycin, a fluorescent analogue of the ionophore valinomycin was synthesized and incorporated into black lipid membranes. Its concentration inside the membrane was measured fluorometrically and was also determined from electrical relaxation experiments, which were analyzed on the basis of a previously proposed carrier model. The results of both methods agreed within less than one order of magnitude. This appears satisfactory in view of the sources of error inherent in both procedures. A conductance increment per carrier molecule of about 3 - 10(-17) omega-1 was obtained for dansyllysine-valinomycin in diphytanoyllecithin membranes at 25 degrees C and 1 M RbCl in the aqueous phases. This is about 400 times smaller compared to unmodified valinomycin in monoolein membranes. The difference is mainly caused by the change in the membrane properties and to a smaller extent by the structural modification of the carrier.

Electric Conductivity↗
PubMedDate not supplied

An extended kinetic analysis of valinomycin-induced Rb-transport through monoglyceride membranes.

The time course of the current following a voltage jump, which is applied to monoglyceride bilayers in the presence of valinomycin, shows two relaxation times. This is basically in agreement with a simple carrier model which has been described in full detail formerly. Relaxation times and amplitudes allow a calculation of the rate constants of the transport model. The presented data supplement an analysis which was hitherto based only on the slower relaxation process and on information derived from the nonlinearity of current-voltage characteristics. The additional resolution of the faster relaxation time allowed an approximate determination of the voltage dependence of the translocation rate constant for carrier-ion-complex and provided evidence for a small voltage dependence of the interfacial reaction. The dependence of the relaxation parameters on the ion concentration in the aqueous phase was interpreted assuming a saturation of the ion concentration at the reaction plane at high bulk concentrations.

Biological Transport↗