Binding and mobility of anti-dinitrophenyl monoclonal antibodies on fluid-like, Langmuir-Blodgett phospholipid monolayers containing dinitrophenyl-conjugated phospholipids

Timbs, Melanie M.; Poglitsch, Claudia L.; Pisarchick, Mary Lee; Sumner, Martina T.; Thompson, Nancy L.

doi:10.1016/0005-2736(91)90305-r

Cited by 26 publications

(13 citation statements)

References 41 publications

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“…A possible physical explanation for the result that cross-linking FL-DPPE with antifluorescein antibodies results in translocation to the raft-phase regions is that protein binding orders the FL-DPPE acyl chains. This notion is consistent with previous work showing that protein binding to lipids in phospholipid monolayers and bilayers can decrease hydrocarbon chain flexibility (49) and reduce lipid translation mobility (50,51). An additional factor might be the increased ability of the larger head groups with their bound antibody to shield cholesterol from water (52).…”

Section: Selective Partitioning Of Cross-linked Components Into Raft supporting

confidence: 92%

Partitioning of Thy-1, GM1, and cross-linked phospholipid analogs into lipid rafts reconstituted in supported model membrane monolayers

Dietrich

Volovyk

Levi

et al. 2001

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

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317

257

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As shown earlier, raft-like domains resembling those thought to be present in natural cell membranes can be formed in supported planar lipid monolayers. These liquid-ordered domains coexist with a liquid-disordered phase and form in monolayers prepared both from synthetic lipid mixtures and lipid extracts of the brush border membrane of mouse kidney cells. The domains are detergent-resistant and are highly enriched in the glycosphingolipid GM1. In this work, the properties of these raft-like domains are further explored and compared with properties thought to be central to raft function in plasma membranes. First, it is shown that domain formation and disruption critically depends on the cholesterol density and can be controlled reversibly by treating the monolayers with the cholesterol-sequestering reagent methyl-␤-cyclodextrin. Second, the glycosylphosphatidylinositol-anchored cell-surface protein Thy-1 significantly partitions into the raft-like domains. The extent of this partitioning is reduced when the monolayers contain GM1, indicating that different molecules can compete for domain occupation. Third, the partitioning of a saturated phospholipid analog into the raft phase is dramatically increased (15% to 65%) after cross-linking with antibodies, whereas the distribution of a doubly unsaturated phospholipid analog is not significantly affected by cross-linking (Ϸ10%). This result demonstrates that cross-linking, a process known to be important for certain cell-signaling processes, can selectively translocate molecules to liquid-ordered domains.membrane domains ͉ receptor cross-linking ͉ cholesterol ͉ glycosylphosphatidylinositol-anchored proteins ͉ signal transduction T reatment of cell lysates with cold nonionic detergent and subsequent sucrose gradient centrifugation allows extraction of a detergent-resistant membrane fraction (DRM) (1, 2). The finding that this fraction is enriched in cholesterol and sphingolipids that form a liquid-ordered phase (3, 4) has led to the hypothesis that the DRM arises directly from discrete liquidordered domains in the plasma membrane termed lipid rafts (5, 6). Such domains could be important for membrane trafficking and sorting (1, 7). Moreover, cell signaling molecules are enriched in the DRM, and the partitioning of these molecules into the DRM is altered during signaling (8-12). Extraction of cholesterol from plasma membranes affects the abundance of molecules found in the DRM and impairs processes like membrane trafficking (13, 14) and cell activation (15, 16). These findings suggest that lipid rafts on plasma membranes are maintained by proper cholesterol levels as functional, preassembled signal transduction complexes (17)(18)(19).Although there is accumulating evidence suggesting that some form of lipid domains or clusters are present in cell membranes, there is still considerable controversy surrounding the basic physical properties of these domains (compositional diversity, size, structure, and dynamics) and their relation to the DRM (for review see refs. 20 and...

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Section: Selective Partitioning Of Cross-linked Components Into Raft supporting

confidence: 92%

Partitioning of Thy-1, GM1, and cross-linked phospholipid analogs into lipid rafts reconstituted in supported model membrane monolayers

Dietrich

Volovyk

Levi

et al. 2001

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

Self Cite

317

257

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“…Lipids were spread at 100 A2/molecule on the air/ water interface of a Joyce-Loebl Langmuir trough (model 4; Vickers Instruments, Inc., Malden, MA) at room temperature (21-250C). The monolayers were compressed at 1-3 A2/molecule per min to a final pressure of 35 dyn/cm and deposited on fused silica microscope slides pretreated with octadecyltrichlorosilane (Aldrich Chemical Co., Milwaukee, IL) by vertical dipping at z5 mm/min (Timbs et al, 1991).…”

Section: Supported Phospholipid Monolayersmentioning

confidence: 99%

“…Previous work has shown that anti-dinitrophenyl (DNP) monoclonal antibodies can bind to these planar model membranes if they contain DNP-conjugated phospholipids. The ability of anti-DNP antibodies to bind to the membrane surfaces and the subsequent spatial arrangements and lateral and rotational mobilities of the bound antibodies are sensitive functions of the physical and chemical properties of the membranes (e.g., Piepenstock and Losche, 1991;Timbs et al, 1991; Pisarchick and Tamm, 1988;Subramaniam et al, 1986;Kimura et al, 1986;Uzgiris and Kornberg, 1983).…”

Section: Introductionmentioning

confidence: 99%

Binding kinetics of an anti-dinitrophenyl monoclonal Fab on supported phospholipid monolayers measured by total internal reflection with fluorescence photobleaching recovery

Pisarchick

Gesty

Thompson

1992

Biophysical Journal

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Fluorescence photobleaching recovery with total internal reflection illumination (TIR-FPR) has been used to measure the dissociation kinetics of a fluorescein-labeled anti-dinitrophenyl monoclonal Fab specifically bound to supported monolayers composed of a mixture of dipalmitoylphosphatidylcholine and dinitrophenyl-conjugated dipalmitoylphosphatidylethanolamine. The fluorescence recovery curves were not monoexponential; when analyzed as a sum of two exponentials, the rates and fractional recoveries were approximately 1 s-1 (approximately 50%) and approximately 0.1 s-1 (approximately 30%). The data did not change as a function of the Fab solution concentration, indicating that the fluorescence recovery curves were not influenced by the rate of diffusion in bulk solution. Also, the recovery curves were independent of the size of the illuminated area, indicating that surface diffusion did not significantly contribute to the rate and shape of the fluorescence recovery. The measured off rates and apparent association constant (1.6 x 10(5) M-1) were analyzed with the theoretical formalism for a proposed mechanism that accounts for the nonmonoexponential kinetics.

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“…Namely, with small immobilized receptor molecules, long, flexible spacers are required to access the deep binding pockets of the proteins that approach the surface. In addition to the spacer length, the efficacy of receptor presentation also reportedly depends on the spacer mobility, surface density, and solvent compatibility (Ebato et al, 1994;Balakrishnan et al, 1982;Tamm & Bartholdus, 1988; Timbs et al, 1991;McConnell et al, 1986;Ahlers et al, 1992). In many cases, the reported surface binding affinities have been significantly lower than those measured in solution.…”

mentioning

confidence: 99%

“…In many cases, the reported surface binding affinities have been significantly lower than those measured in solution. The latter discrepancies have been attributed to the surface microenvironment (e.g., local pH) (McLaughlin, 1989), to mobility losses (Ebato et al, 1994), to large ligands or receptors-that is, to twodimensional nonideal excluded area effects (Stankowski, 1983), and to steric hindrance at high receptor surface densities (Tamm & Bartholdus, 1988;Timbs et al, 1991).…”

mentioning

confidence: 99%

4-4-20 Anti-Fluorescyl IgG Fab' Recognition of Membrane Bound Hapten: Direct Evidence for the Role of Protein and Interfacial Structure

Kuhl

et al. 1995

Biochemistry

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The surface forces apparatus was used to identify the molecular forces that control the interactions of monoclonal 4-4-20 antifluorescyl IgG Fab' fragments with fluorescein-presenting supported planar bilayers. At long range, the electrostatic force between oriented Fab' and fluorescein monolayers was controlled by the composition of the protein exterior surrounding the antigen-combining site rather than by the overall protein charge. The measured positive electrostatic potential of the Fab' monolayer at pH > pI(Fab') was consistent with the structure of the exposed Fab' surface in which a ring of positive charge at the mouth of the antigen-combining site dominates the local electrostatic surface properties. Substantial differences in the electrostatic forces measured with denatured Fab' further demonstrated that the measured electrostatic surface properties and the consequent long-range interaction forces are controlled by the protein surface composition. At short range, the strength of the Fab'-mediated adhesion was modulated not only by the length of the fluorescein tether but also by membrane hydration. Steric hydration barriers at the membrane surface reduced the adhesion strength in proportion to their range of influence. These results provide direct evidence that long-range protein interactions with immobilized ligands are controlled by both the protein and the membrane surface compositions, while short-range, specific binding is modulated by both the protein structure and the membrane interfacial properties.

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Binding and mobility of anti-dinitrophenyl monoclonal antibodies on fluid-like, Langmuir-Blodgett phospholipid monolayers containing dinitrophenyl-conjugated phospholipids

Cited by 26 publications

References 41 publications

Partitioning of Thy-1, GM1, and cross-linked phospholipid analogs into lipid rafts reconstituted in supported model membrane monolayers

Partitioning of Thy-1, GM1, and cross-linked phospholipid analogs into lipid rafts reconstituted in supported model membrane monolayers

Binding kinetics of an anti-dinitrophenyl monoclonal Fab on supported phospholipid monolayers measured by total internal reflection with fluorescence photobleaching recovery

4-4-20 Anti-Fluorescyl IgG Fab' Recognition of Membrane Bound Hapten: Direct Evidence for the Role of Protein and Interfacial Structure

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