0000000000165147
AUTHOR
Deborah E. Leckband
showing 5 related works from this author
4-4-20 anti-fluorescyl IgG Fab' recognition of membrane bound hapten: direct evidence for the role of protein and interfacial structure.
1995
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 s…
Force probe measurements of antibody-antigen interactions.
2000
The surface force apparatus has been used to quantify directly the forces that govern the interactions between proteins and ligands. In this work, we describe the measured interactions between the antigen fluorescein and the Fab' fragment of the monoclonal 4-4-20 anti-fluorescyl IgG antibody. Here we first describe the use of the surface force apparatus to demonstrate directly the impact of the charge composition in the region of the antibody binding site on the antibody interactions. Several approaches are described for immobilizing antigens, antibodies, and proteins in general for direct force measurements. The measured force profiles presented are accompanied by an extensive discussion o…
Molecular mechanisms determining the strength of receptor-mediated intermembrane adhesion
1995
The strength of receptor-mediated cell adhesion is directly controlled by the mechanism of cohesive failure between the cell surface and underlying substrate. Unbinding can occur either at the locus of the specific bond or within the bilayer, which results in tearing the hydrophobic anchors from the membrane interior. In this work, the surface force apparatus has been used to investigate the relationship between the receptor-ligand bond affinities and the dominant mechanism of receptor-coupled membrane detachment. The receptors and ligands used in this study were membrane-bound streptavidin and biotin analogs, respectively, with solution affinities ranging over 10 orders of magnitude. With …
Role of calcium in the adhesion and fusion of bilayers.
1993
The interaction forces and fusion mechanisms of mixed zwitterionic-anionic phospholipid bilayers were measured with the surface forces apparatus. The bilayers were 3:1 mixtures of either dimyristoylphosphatidylcholine and dimyristoylphosphatidylglycerol (DMPC/DMPG) or dilauroylphosphatidylcholine and dilauroylphosphatidylglycerol (DLPC/DLPG), and experiments were carried out in NaCl solutions with and without CaCl2. In NaCl solutions, the forces between either mixed bilayer system were consistent with the DLVO (Derjaguin-Landau-Verwey-Overbeek) theory of repulsive electrostatic and attractive van der Waals forces, and fusion did not occur. At high pH (> 6) and in high (20 mM) NaCl concentra…
Specific Protein Binding to Functionalized Interfaces
1992
We report on the characterization of specific binding reactions between streptavidin and biotinylated model membrane surfaces. Self-assembly techniques as well as the Langmuir-Blodgett-Kuhn method were employed to prepare reactive, functionalized surfaces on various solid supports in contact with the aqueous protein solution. Plasmon surface polaritons optical measurements as well as atomic force microscopy and studies with the surface forces apparatus give rather detailed information as to the streptavidin monolayer formation, the kinetics of this process (either binding site- or diffusion limited), the selectivity of the reaction at laterally heterogeneous membranes, and the involved inte…