Location of Repository

Structural requirements for adhesion of soluble recombinant murine vascular cell adhesion molecule-1 to alpha 4 beta 1



This study describes the identification of seven amino acid residues of the vascular cell adhesion molecule (VCAM-1) that influence binding to the alpha 4 beta 1 receptor. Using recombinant murine VCAM-1-IgG, which is bound by both mouse (WEHI 231) and human (Ramos) lymphoid cells, two approaches demonstrated the crucial role of the first two NH2-terminal Ig-like domains in binding: (a) blocking monoclonal anti-mouse VCAM-1 antibodies bound to only truncation variants that included the first two domains; (b) site-direct mutagenesis of the first NH2-terminal domain showed that alanine substitution of the amino acid residues R36, D40, K46, S54, N65, T72, and E81 partially or completely reduced adherence by human and/or mouse cells. Of these D40, when mutated to A, N, or K (but not E), showed complete abrogation of adherence by mouse and human cells, as well as inability to bind blocking anti-murine VCAM- 1 antibody MVCAM.A429, while not inducing gross structural perturbations in VCAM-1. By molecular modeling, the D40 residue was located on a beta turn connecting two beta strands defined as C and D. The residues R36, K46, S54, N65, T72, and E81, which perturb cell adherence and caused small changes to gross structure, are conformationally near or adjacent to D40. Although these residues, identified as crucial for cell adhesion, are all located in domain 1, it is evident that there is a structural requirement for domains 1 and 2 to be intact so that cell adhesive function can occur

Topics: Articles
Publisher: The Rockefeller University Press
OAI identifier: oai:pubmedcentral.nih.gov:2290915
Provided by: PubMed Central
Download PDF:
Sorry, we are unable to provide the full text but you may find it at the following location(s):
  • http://www.pubmedcentral.nih.g... (external link)
  • Suggested articles



    1. (1990). A homing receptor IgG chimera as a probe for adhesive ligands of lymph node high endothelial vennles.
    2. (1993). A monoclonal antibody recognizing vary late activation antigen-4 inhibits ¢osinophil accumulation in vivo.
    3. (1991). A VCAM-like adhesion molecule on routine bone marrow stromal cells mediates binding of lymphocyte precursors in culture.
    4. (1992). Activated endotbelium binds lymphocytes through a novel binding site in the alternately spliced domain of vascular cell cell adhesion molecule-1.
    5. (1992). Adhesion to vascular cell adhesion molecule 1 and fibronectin.
    6. (1986). An all atom force field for simulation of proteins and nucleic acids.
    7. (1993). Antigen-independent processes in antigen-specific immunity.
    8. (1991). Cloning of an alternative form of vascular cell adhesion molecule-1 (VCAM-1).
    9. (1993). Cloning of an inflammation-specific phosphatidyl inositol-liuked form of murine vascular cell adhesion molecule-1.
    10. (1992). Cloning of murine and rat vascular cell adhesion molecule-1.
    11. (1991). Differential utilization of ICAM-1 and VCAM-1 during the adhesion and ttansendothelial migration of human T lymphocytes.
    12. (1989). Endothelial interleukin 8: a novel inhibitor of leukocyte-endothelial interaction.
    13. (1991). Evidence for a role of the integrin VLA-4 in lympho-hemopoiesis.
    14. (1992). Expression of adhesion molecules in human intestinal graft-versus-host disease.
    15. (1991). Expression of VCAM-1 in normal and diseased kidney.
    16. (1991). Four molecular pathways of T cell adhesion to endothelial cells: roles of LFA-1, VCAM-1, and ELAM-1 and changes in pathway hierarchy under different activation conditions.
    17. (1990). Full length vascular cell adhesion molecule 1 (VCAM-1). Nucleic Acids Res.
    18. (1991). Functional evidence for three distinct and independently inhibitable adhesion activities mediated by the human integrin VLA-4. Correlation with distinct or4 epitopes.
    19. (1991). G-ene structure, chromosome location, and basis for alternative mRNA splicing of the human VCAMI gene.
    20. (1993). Homotypic leukocyte aggregation triggered by a monoclonal antibody specific for a novel epitope expressed by the integrin/31 subunit: conversion of nonrespousive cells by transfecting human integrin c~4 subunit cDNA.
    21. (1991). Human monocytes bind to two cytokine-induced adhesive ligands on cultured human endothelial cells: endothelial-leukocyte adhesion molecules-I and vascular cell adhesion molecule-1.
    22. (1991). Immunolocalization of endothelial and leukocyte adhesion molecules in human rheumatoid and osteoarthritic synovial tissues.
    23. (1993). Implications of de novo ELAM-1 and VCAM-1 expression in human cardiac allograft rejection. Transplammion
    24. (1992). Importance of endothelial VCAM-1 for inflammatory leukocytic infiltration in vivo.
    25. (1992). Increased binding of synovial T lymphocytes from rheumatoid arthritis to endothelial-leukocyte adhesion molecule-1 (ELAM-1) and vascular cell adhesion molecule-1 (VCAM-1).
    26. (1991). Induced expression of endothelial leukocyte adhesion molecules in human cardiac allograRs. Transplantation
    27. (1990). Inducible cell adhesion molecule 110 0NCAM-110) is an endothelial receptor for lymphocytes. A CD11/CD18-independent adhesion mechanism.
    28. (1992). Induction of VCAM-1 and ICAM-1 on human neural cells and mechanisms of mononuclear leukocyte adherence.
    29. (1992). Lymphocyte adhesion through very late antigen 4: evidence for a novel binding site in the alternatively spliced domain of vascular cell adhesion molecule I and an additional ct4 integrin counter-receptor on stimulated endothalium.
    30. (1993). Monoclonal antibody to anti-VCAM-I interferes with murine cardiac allograft rejection.
    31. (1992). Prevention of experimental autoimmune encephalomyelitis by antibodies against t~4/31 integrin.
    32. (1987). Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol.
    33. (1992). Role of integrin o~4ff'//a4/3P in lymphocyte adherence to fibronectin and VCAM-1 and in homotypic cell clustering.
    34. (1992). Roles for the integrin VLA-4 and its counter receptor VCAM-I in myogenesis.
    35. (1991). Structural and functional studies of full-length vascular cell adhesion molecule-l: Internal duplication and homology to several adhesion proteins.
    36. (1988). Structure and function of human and murine receptors for IgG.
    37. Structure/function studies on vascular cell adhesion molecule-1.
    38. Surface expression of 64 integrin by CD4 T cells is required for their entry into brain pareuchyma.
    39. (1987). Tertiary template for proteins of packing criteria in the numeration of allowed sequences for different structural classes.
    40. The protein databank: a computer-based archival file for macromolecular studies.
    41. (1991). The VLA4/VCAM-1 pathway is involved in lymphocyte adhesion to endothelinm in rheumatoid synovium.
    42. (1991). Tumor necrosis factor combines with IL-4 or IFN-? to selectively enhance endothelial cell adhesiveness for T cells. The contribution of vascular cell adhesion molecule-1-dependent and -independent binding mechanisms.
    43. (1991). Vascular and non-vascular expression of INCAM-110. A target for mononuclear leukocyte adhesion in normal and inflamed human tissues.
    44. (1991). Vascular cell adhesion molecule 1 induces T-cell antigen receptor-dependent activation of CD4 ÷ T ]ymphocytes.
    45. (1991). Vascular cell adhesion molecule-1 and the iutegrin VIA-4 mediate adhesion of human cell precursors to cultured bone marrow adherent cells.
    46. (1990). Vascular cell adhesion molecule-1 mediates lymphocyte adhesion to cytokine-activated cultured human endothelial cells.
    47. (1992). Vascular cell adhesion molecules-I (VCAM-1) expression in routine lupus nephritis.
    48. (1990). VCAM-1 on activated endothelium interact with the leukocyte integrin VLA-4 at a site distinct from the VLA4/fibronectin binding site.
    49. (1986). VLA-3: a novel polypeptide association within the VLA molecular complex: cell distribution and biochemical characterization.
    50. X-ray structures of the antigen-binding domains from three variants of humanized anti-p185HER2 antibody 4D5 and comparison with molecular modeling.

    To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.