Differential binding of VEGF isoforms to VEGF receptor 2 in the presence of neuropilin-1: a computational model.

A comprehensive, biophysically accurate, computational model of vascular endothelial growth factor (VEGF) family member interactions with endothelial cell surface receptors was developed to study angiogenesis. Neuropilin-1 (NRP1) and the signaling VEGF receptor, VEGFR2, do not interact directly but are bridged by one VEGF isoform, VEGF(165). Using the model and published experimental data, we estimated the kinetic rate of this VEGFR2-NRP1 coupling in vitro. With the use of this rate, our model gives predictions in good quantitative agreement with several independent in vitro experiments involving VEGF(121) and VEGF(165) isoforms, confirming that VEGFR2-NRP1 coupling through VEGF(165) can fully explain the observed differences in receptor binding and phosphorylation in response to these isoforms. Model predictions also determine the mechanism of action of a commonly used NRP1 antibody and predict the results of potential future experiments. This is the first model to include VEGF isoforms or NRPs, and it is a necessary step toward a quantitative molecular level description of VEGF that can be extended to in vivo situations. The model has applications for both proangiogenic and antiangiogenic therapies, such as for heart disease and cancer, as well as in tissue engineering.

[1]  M. Tessier-Lavigne,et al.  Neuropilins as Semaphorin Receptors , 2002 .

[2]  D A Lauffenburger,et al.  The role of low-affinity interleukin-2 receptors in autocrine ligand binding: alternative mechanisms for enhanced binding effect. , 1994, Molecular immunology.

[3]  M. Tessier-Lavigne,et al.  Class 3 semaphorins control vascular morphogenesis by inhibiting integrin function , 2003, Nature.

[4]  S. Cunningham,et al.  Characterization of Vascular Endothelial Cell Growth Factor Interactions with the Kinase Insert Domain-containing Receptor Tyrosine Kinase , 1999, The Journal of Biological Chemistry.

[5]  D. Jans.  The mobile receptor hypothesis revisited: a mechanistic role for hormone receptor lateral mobility in signal transduction. , 1992, Biochimica et biophysica acta.

[6]  Aleksander S. Popel,et al.  A Reaction-Diffusion Model of Basic Fibroblast Growth Factor Interactions with Cell Surface Receptors , 2004, Annals of Biomedical Engineering.

[7]  B. J. Limberg,et al.  Vascular Endothelial Growth Factor Receptor-2 and Neuropilin-1 Form a Receptor Complex That Is Responsible for the Differential Signaling Potency of VEGF165 and VEGF121 * , 2001, The Journal of Biological Chemistry.

[8]  Nijole Gasiunas,et al.  Neuropilin-1 Binds Vascular Endothelial Growth Factor 165, Placenta Growth Factor-2, and Heparin via Its b1b2 Domain* , 2002, The Journal of Biological Chemistry.

[9]  M. Backer,et al.  Kinase Insert Domain Receptor (KDR) Extracellular Immunoglobulin-like Domains 4–7 Contain Structural Features That Block Receptor Dimerization and Vascular Endothelial Growth Factor-induced Signaling* , 2001, The Journal of Biological Chemistry.

[10]  VEGF receptor signal transduction. , 2003, Science's STKE : signal transduction knowledge environment.

[11]  Shinji Koyama,et al.  Selective induction of neuropilin-1 by vascular endothelial growth factor (VEGF): A mechanism contributing to VEGF-induced angiogenesis , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[12]  T. Yagi,et al.  A requirement for neuropilin-1 in embryonic vessel formation. , 1999, Development.

[13]  T. Kitsukawa,et al.  Targeting of both mouse neuropilin-1 and neuropilin-2 genes severely impairs developmental yolk sac and embryonic angiogenesis , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[14]  K. Garcia,et al.  The interaction of neuropilin-1 with vascular endothelial growth factor and its receptor flt-1. , 2000, The Journal of biological chemistry.

[15]  B. Keyt,et al.  Identification of Vascular Endothelial Growth Factor Determinants for Binding KDR and FLT-1 Receptors , 1996, The Journal of Biological Chemistry.

[16]  U. Hellman,et al.  Differential Binding of Vascular Endothelial Growth Factor B Splice and Proteolytic Isoforms to Neuropilin-1* , 1999, The Journal of Biological Chemistry.

[17]  P E Thorpe,et al.  Expression of soluble VEGF receptor 2 and characterization of its binding by surface plasmon resonance. , 1998, Biochemical and biophysical research communications.

[18]  J. Linderman,et al.  Calculation of diffusion-limited kinetics for the reactions in collision coupling and receptor cross-linking. , 1997, Biophysical journal.

[19]  T. L. Hill,et al.  Free Energy Transduction in Biology: The Steady-State Kinetic and Thermodynamic Formalism , 1977 .

[20]  Shay Soker,et al.  VEGF165 mediates formation of complexes containing VEGFR‐2 and neuropilin‐1 that enhance VEGF165‐receptor binding , 2002, Journal of cellular biochemistry.

[21]  Bernd Fritzsch,et al.  Neuropilin-1 conveys semaphorin and VEGF signaling during neural and cardiovascular development. , 2003, Developmental cell.

[22]  Shay Soker,et al.  Characterization of Novel Vascular Endothelial Growth Factor (VEGF) Receptors on Tumor Cells That Bind VEGF via Its Exon 7-encoded Domain (*) , 1996, The Journal of Biological Chemistry.

[23]  A. Popel,et al.  Model of competitive binding of vascular endothelial growth factor and placental growth factor to VEGF receptors on endothelial cells. , 2004, American journal of physiology. Heart and circulatory physiology.

[24]  N Almqvist,et al.  Elasticity and adhesion force mapping reveals real-time clustering of growth factor receptors and associated changes in local cellular rheological properties. , 2004, Biophysical journal.

[25]  M. Kan,et al.  Vascular endothelial cell growth factor (VEGF) produced by A-431 human epidermoid carcinoma cells and identification of VEGF membrane binding sites. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[26]  L. Aiello,et al.  Comparative Analysis of Vascular Endothelial Growth Factor Receptors on Retinal and Aortic Vascular Endothelial Cells , 1995, Diabetes.

[27]  T. Kitsukawa,et al.  Overexpression of a membrane protein, neuropilin, in chimeric mice causes anomalies in the cardiovascular system, nervous system and limbs. , 1995, Development.

[28]  Shay Soker,et al.  Neuropilin-1 Is Expressed by Endothelial and Tumor Cells as an Isoform-Specific Receptor for Vascular Endothelial Growth Factor , 1998, Cell.

[29]  Hung V. Nguyen,et al.  The Carboxyl-terminal Domain(111165) of Vascular Endothelial Growth Factor Is Critical for Its Mitogenic Potency (*) , 1996, The Journal of Biological Chemistry.

[30]  M. Dembo,et al.  Kinetic analysis of histamine release due to covalently linked IgE dimers. , 1982, Molecular immunology.

[31]  B Z Levi,et al.  VEGF121, a vascular endothelial growth factor (VEGF) isoform lacking heparin binding ability, requires cell-surface heparan sulfates for efficient binding to the VEGF receptors of human melanoma cells , 1995, The Journal of Biological Chemistry.

[32]  R K Jain,et al.  Fluorescence photobleaching with spatial Fourier analysis: measurement of diffusion in light-scattering media. , 1993, Biophysical journal.

[33]  D. Mukhopadhyay,et al.  Neuropilin-1-mediated Vascular Permeability Factor/Vascular Endothelial Growth Factor-dependent Endothelial Cell Migration* , 2003, Journal of Biological Chemistry.

[34]  M. Shibuya,et al.  Neuropilin-1 Is a Placenta Growth Factor-2 Receptor* , 1998, The Journal of Biological Chemistry.

[35]  M. Shibuya,et al.  MMP9 induction by vascular endothelial growth factor receptor-1 is involved in lung-specific metastasis. , 2002, Cancer cell.

[36]  Shay Soker,et al.  Relative effects of VEGF‐A and VEGF‐C on endothelial cell proliferation, migration and PAF synthesis: Role of neuropilin‐1 , 2002, Journal of cellular biochemistry.

[37]  Chenghua Gu,et al.  Characterization of Neuropilin-1 Structural Features That Confer Binding to Semaphorin 3A and Vascular Endothelial Growth Factor 165* , 2002, The Journal of Biological Chemistry.

[38]  I. Zachary,et al.  Signaling transduction mechanisms mediating biological actions of the vascular endothelial growth factor family. , 2001, Cardiovascular research.

[39]  Alex L Kolodkin,et al.  Neuropilin Is a Semaphorin III Receptor , 1997, Cell.

[40]  Alex L Kolodkin,et al.  Neuropilin-2 Is a Receptor for Semaphorin IV Insight into the Structural Basis of Receptor Function and Specificity , 1998, Neuron.

[41]  M. Bryckaert,et al.  Interaction of vasculotropin/vascular endothelial cell growth factor with human umbilical vein endothelial cells: Binding, internalization, degradation, and biological effects , 1991, Journal of cellular physiology.

[42]  P. Valent,et al.  Characterization of 123I‐vascular endothelial growth factor–binding sites expressed on human tumour cells: Possible implication for tumour scintigraphy , 2001, International journal of cancer.

[43]  M. Tessier-Lavigne,et al.  Neuropilins as Semaphorin receptors: in vivo functions in neuronal cell migration and axon guidance. , 2002, Advances in experimental medicine and biology.

[44]  M. Shibuya,et al.  Different signal transduction properties of KDR and Flt1, two receptors for vascular endothelial growth factor. , 1994, The Journal of biological chemistry.