Strength of multiple parallel biological bonds.

Multivalent interactions play a critical role in a variety of biological processes on both molecular and cellular levels. We have used molecular force spectroscopy to investigate the strength of multiple parallel peptide-antibody bonds using a system that allowed us to determine the rupture forces and the number of ruptured bonds independently. In our experiments the interacting molecules were attached to the surfaces of the probe and sample of the atomic force microscope with flexible polymer tethers, and the unique mechanical signatures of the tethers determined the number of ruptured bonds. We show that the rupture forces increase with the number of interacting molecules and that the measured forces obey the predictions of a Markovian model for the strength of multiple parallel bonds. We also discuss the implications of our results to the interpretation of force spectroscopy measurements in multiple bond systems.

[1]  M. Rief,et al.  Reversible unfolding of individual titin immunoglobulin domains by AFM. , 1997, Science.

[2]  Matthias Rief,et al.  Single molecule force spectroscopy by AFM indicates helical structure of poly(ethylene-glycol) in water , 1999 .

[3]  Aleksandr Noy,et al.  Dynamic force spectroscopy of parallel individual Mucin1-antibody bonds. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[4]  Alexander Miller,et al.  Antigen-Specific Signaling by a Soluble, Dimeric Peptide/Major Histocompatibility Complex Class II/Fc Chimera Leading to T Helper Cell Type 2 Differentiation , 1999, The Journal of experimental medicine.

[5]  Hendrik Dietz,et al.  Exploring the energy landscape of GFP by single-molecule mechanical experiments. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[6]  G. Denardo,et al.  Production of Soluble ScFvs with C-Terminal-Free Thiol for Site-Specific Conjugation or Stable Dimeric ScFvs on Demand , 2004 .

[7]  C. MacKenzie,et al.  Analysis by Surface Plasmon Resonance of the Influence of Valence on the Ligand Binding Affinity and Kinetics of an Anti-carbohydrate Antibody (*) , 1996, The Journal of Biological Chemistry.

[8]  M. Davies,et al.  Influence of architecture on the kinetic stability of molecular assemblies. , 2004, Journal of the American Chemical Society.

[9]  E. Evans,et al.  Dynamic strength of molecular adhesion bonds. , 1997, Biophysical journal.

[10]  T. Beebe,et al.  Determination of Single-Bond Forces from Contact Force Variances in Atomic Force Microscopy , 1996 .

[11]  M. Ho,et al.  Synergism of multiple adhesion molecules in mediating cytoadherence of Plasmodium falciparum–infected erythrocytes to microvascular endothelial cells under flow , 2000 .

[12]  R. Merkel,et al.  Energy landscapes of receptor–ligand bonds explored with dynamic force spectroscopy , 1999, Nature.

[13]  M. J. Mattes,et al.  Re-evaluation of the concept of functional affinity as applied to bivalent antibody binding to cell surface antigens. , 1993, Molecular immunology.

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

[15]  H. Güntherodt,et al.  Dynamic force spectroscopy of single DNA molecules. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[16]  P. Underwood.  Problems and pitfalls with measurement of antibody affinity using solid phase binding in the ELISA. , 1993, Journal of immunological methods.

[17]  H. Gaub,et al.  Adhesion forces between individual ligand-receptor pairs. , 1994, Science.

[18]  H. Flyvbjerg,et al.  Physics of bio-molecules and cells , 2002 .

[19]  P. Williams.  Analytical descriptions of dynamic force spectroscopy: behaviour of multiple connections , 2003 .

[20]  R. Webster,et al.  Single-chain Fv fragments of anti-neuraminidase antibody NC10 containing five- and ten-residue linkers form dimers and with zero-residue linker a trimer. , 1997, Protein engineering.

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

[22]  T Prospero,et al.  "Diabodies": small bivalent and bispecific antibody fragments. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[23]  S. Denardo,et al.  Systemic radiotherapy in metastatic breast cancer using 90Y-linked monoclonal MUC-1 antibodies. , 2001, Critical reviews in oncology/hematology.

[24]  Jean-Jacques Meister,et al.  Short-term binding of fibroblasts to fibronectin: optical tweezers experiments and probabilistic analysis , 2000, European Biophysics Journal.