Quantification of cell adhesion force with AFM: distribution of vitronectin receptors on a living MC3T3-E1 cell.

Distribution of vitronectin (VN) receptors on a living murine osteoblastic cell was successfully measured by atomic force microscopy (AFM). First, the distribution of the integrin beta(5) subunit which constitutes a part of the VN receptor on the cell was confirmed by conventional immunohistochemistry after fixing the cell. To visualize the distribution of the receptor on a living cell by an independent and potentially a more quantitative method, the AFM was used with a microbead attached to the cantilever tip to increase the area of contact and VN was immobilized on the microbead. Force measurements were then performed over a large area of a living murine osteoblastic cell using the microbead covered with VN.

[1]  S. Jarvis,et al.  Limits of imaging resolution for atomic force microscopy of molecules , 1991 .

[2]  E. Wayner,et al.  Integrins alpha v beta 3 and alpha v beta 5 contribute to cell attachment to vitronectin but differentially distribute on the cell surface , 1991, The Journal of cell biology.

[3]  A. Ikai,et al.  Force measurement and inhibitor binding assay of monomer and engineered dimer of bovine carbonic anhydrase B. , 2001, Biochemical and biophysical research communications.

[4]  M. Ichikawa,et al.  The distribution of sugar chains on the vomeronasal epithelium observed with an atomic force microscope. , 1999, Chemical senses.

[5]  M A Horton,et al.  Single integrin molecule adhesion forces in intact cells measured by atomic force microscopy. , 1999, Biochemical and biophysical research communications.

[6]  D. Seiffert,et al.  Role of vitronectin and its receptors in haemostasis and vascular remodeling. , 1998, Thrombosis research.

[7]  D. Woodley,et al.  Vitronectin-driven human keratinocyte locomotion is mediated by the alpha v beta 5 integrin receptor. , 1994, The Journal of biological chemistry.

[8]  Jeffrey W. Smith,et al.  Divalent cations regulate the organization of integrins αvβ3 and αvβ5 on the cell surface , 1996 .

[9]  Atsushi Ikai,et al.  Quantification of fibronectin and cell surface interactions by AFM , 2002 .

[10]  Richard O. Hynes,et al.  Integrins: Versatility, modulation, and signaling in cell adhesion , 1992, Cell.

[11]  I. N. Sneddon.  The relation between load and penetration in the axisymmetric boussinesq problem for a punch of arbitrary profile , 1965 .

[12]  J. Depasquale.  Cell matrix adhesions and localization of the vitronectin receptor in MCF-7 human mammary carcinoma cells , 1998, Histochemistry and Cell Biology.

[13]  H. Gaub,et al.  Intermolecular forces and energies between ligands and receptors. , 1994, Science.

[14]  E Ruoslahti,et al.  New perspectives in cell adhesion: RGD and integrins. , 1987, Science.

[15]  V. Moy,et al.  Cross-linking of cell surface receptors enhances cooperativity of molecular adhesion. , 2000, Biophysical journal.

[16]  A Ikai,et al.  Dynamic measurement of single protein's mechanical properties. , 2000, Biochemical and biophysical research communications.

[17]  D. Cheresh,et al.  Purification and functional characterization of integrin alpha v beta 5. An adhesion receptor for vitronectin. , 1990, The Journal of biological chemistry.

[18]  Atsushi Ikai,et al.  STM and AFM of bio/organic molecules and structures , 1996 .

[19]  Hyonchol Kim,et al.  Quantification of cell adhesion interactions by AFM: effects of LPS/PMA on the adhesion of C6 glioma cell to collagen type I , 2002 .