Measuring two-dimensional receptor-ligand binding kinetics by micropipette.

We report a novel method for measuring forward and reverse kinetic rate constants, kf0 and kr0, for the binding of individual receptors and ligands anchored to apposing surfaces in cell adhesion. Not only does the method examine adhesion between a single pair of cells; it also probes predominantly a single receptor-ligand bond. The idea is to quantify the dependence of adhesion probability on contact duration and densities of the receptors and ligands. The experiment was an extension of existing micropipette protocols. The analysis was based on analytical solutions to the probabilistic formulation of kinetics for small systems. This method was applied to examine the interaction between Fc gamma receptor IIIA (CD16A) expressed on Chinese hamster ovary cell transfectants and immunoglobulin G (IgG) of either human or rabbit origin coated on human erythrocytes, which were found to follow a monovalent biomolecular binding mechanism. The measured rate constants are Ackf0 = (2.6 +/- 0.32) x 10(-7) micron 4 s-1 and kr0 = (0.37 +/- 0.055) s-1 for the CD16A-hIgG interaction and Ackf0 = (5.7 +/- 0.31) X 10(-7) micron 4 s-1 and kr0 = (0.20 +/- 0.042) s-1 for the CD16A-rIgG interaction, respectively, where Ac is the contact area, estimated to be a few percent of 3 micron 2.

[1]  Shu Chien,et al.  Handbook of Bioengineering , 1986 .

[2]  S Chien,et al.  Determination of junction avidity of cytolytic T cell and target cell. , 1986, Science.

[3]  E. Evans,et al.  Sensitive force technique to probe molecular adhesion and structural linkages at biological interfaces. , 1995, Biophysical journal.

[4]  T. Springer,et al.  The major Fc receptor in blood has a phosphatidylinositol anchor and is deficient in paroxysmal nocturnal haemoglobinuria , 1988, Nature.

[5]  T. Springer,et al.  Leukocytes roll on a selectin at physiologic flow rates: Distinction from and prerequisite for adhesion through integrins , 1991, Cell.

[6]  H. Fudenberg,et al.  Chromic chloride: a coupling reagent for passive hemagglutination reactions. , 1967, Journal of immunology.

[7]  E. Unanue.  Antigen-presenting function of the macrophage. , 1984, Annual review of immunology.

[8]  O. Coenen,et al.  Interaction forces between red cells agglutinated by antibody. IV. Time and force dependence of break-up. , 1993, Biophysical journal.

[9]  P. Bongrand,et al.  Experimental Study of the Rate of Bond Formation Betwwen Individual Receptor-Coated Spheres and Ligand-Bearing Surfaces , 1996 .

[10]  H Schindler,et al.  Detection and localization of individual antibody-antigen recognition events by atomic force microscopy. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[11]  William H. Press,et al.  Numerical recipes in C. The art of scientific computing , 1987 .

[12]  R. Swerlick,et al.  A CENTRIFUGATION METHOD FOR MEASUREMENT OF TWO-DIMENSIONAL BINDING CHARACTERISTICS OF RECEPTOR-LIGAND INTERACTION , 1999 .

[13]  R. Ghirlando,et al.  Stoichiometry and thermodynamics of the interaction between the Fc fragment of human IgG1 and its low-affinity receptor Fc gamma RIII. , 1995, Biochemistry.

[14]  G. I. Bell.  Models for the specific adhesion of cells to cells. , 1978, Science.

[15]  G. Scatchard,et al.  THE ATTRACTIONS OF PROTEINS FOR SMALL MOLECULES AND IONS , 1949 .

[16]  S. Erlandsen,et al.  Interactions of the Cytoplasmic Domain of P-Selectin with Clathrin-coated Pits Enhance Leukocyte Adhesion under Flow , 1998, The Journal of cell biology.

[17]  Donald A. McQuarrie,et al.  Kinetics of Small Systems. I , 1963 .

[18]  Timothy A. Springer,et al.  The Kinetics of L-selectin Tethers and the Mechanics of Selectin-mediated Rolling , 1997, The Journal of cell biology.

[19]  J D Chambers,et al.  Two-step model of leukocyte-endothelial cell interaction in inflammation: distinct roles for LECAM-1 and the leukocyte beta 2 integrins in vivo. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[20]  P. Anderson,et al.  Ligand binding and phagocytosis by CD16 (Fc gamma receptor III) isoforms. Phagocytic signaling by associated zeta and gamma subunits in Chinese hamster ovary cells. , 1995, The Journal of biological chemistry.

[21]  C. Zhu,et al.  Determining force dependence of two-dimensional receptor-ligand binding affinity by centrifugation. , 1998, Biophysical journal.

[22]  M. Hulett,et al.  Molecular basis of Fc receptor function. , 1994, Advances in immunology.

[23]  E. Evans,et al.  PROBING MOLECULAR ATTACHMENTS TO CELL SURFACE RECEPTORS: Image of Stochastic Bonding and Rupture Processes , 1994 .

[24]  H. Goldsmith,et al.  Kinetics and locus of failure of receptor-ligand-mediated adhesion between latex spheres. I. Protein-carbohydrate bond. , 1996, Biophysical journal.

[25]  W. Bensinger,et al.  Positive selection of viable cell populations using avidin-biotin immunoadsorption. , 1986, Journal of immunological methods.

[26]  P Bongrand,et al.  Measuring the Lifetime of Bonds Made between Surface-linked Molecules (*) , 1995, The Journal of Biological Chemistry.

[27]  T. Springer,et al.  Rolling and transient tethering of leukocytes on antibodies reveal specializations of selectins. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[28]  R. Kofler,et al.  Some methodologic aspects of the chromium chloride method for coupling antigen to erythrocytes. , 1977, Journal of immunological methods.

[29]  P Bongrand,et al.  Measuring bonds between surface-associated molecules. , 1996, Journal of immunological methods.

[30]  S. Simon,et al.  Preservation of spatial organization and antigenicity of leukocyte surface molecules by aldehyde fixation: flow cytometry and high-resolution FESEM studies of CD62L, CD11b, and Thy-1. , 1996, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[31]  D. Hammer,et al.  Lifetime of the P-selectin-carbohydrate bond and its response to tensile force in hydrodynamic flow , 1995, Nature.

[32]  D A Lauffenburger,et al.  Receptor-mediated cell attachment and detachment kinetics. I. Probabilistic model and analysis. , 1990, Biophysical journal.

[33]  A Leung,et al.  Detachment of agglutinin-bonded red blood cells. I. Forces to rupture molecular-point attachments. , 1991, Biophysical journal.

[34]  J. V. D. van de Winkel,et al.  Human IgG Fc receptor heterogeneity: molecular aspects and clinical implications. , 1993, Immunology today.

[35]  D E Leckband,et al.  Direct force measurements of specific and nonspecific protein interactions. , 1994, Biochemistry.

[36]  Michael Loran Dustin,et al.  Visualization of CD2 interaction with LFA-3 and determination of the two-dimensional dissociation constant for adhesion receptors in a contact area , 1996, The Journal of cell biology.

[37]  G. I. Bell,et al.  Concanavalin-A-mediated thymocyte agglutination: a model for a quantitative study of cell adhesion. , 1982, Journal of cell science.

[38]  William H. Press,et al.  Book-Review - Numerical Recipes in Pascal - the Art of Scientific Computing , 1989 .

[39]  S Kaplanski,et al.  Granulocyte-endothelium initial adhesion. Analysis of transient binding events mediated by E-selectin in a laminar shear flow. , 1993, Biophysical journal.