Purification and characterization of scatter factor

  • Ermanno Gherardi
  • Arnold Coffer
Part of the Experientia Supplementum book series (EXS, volume 59)


Scatter factor is a fibroblast-derived protein which disrupts and scatters epithelial colonies and enhances the local movement of individual epithehal and endothehal cells. The factor purified from mouse fibroblasts by cation-exchange and reverse phase chromatography is a dimer of 57 kD and 30 kD protein subunits (A and B subunits), is active at picomolar concentrations and requires intact intra- and/or inter-chain disulphide bonds for activity. In serum-free conditioned medium the factor is highly aggregated but in the presence of high-salt buffers or protein denaturants elutes from gel filtration columns with an apparent Mr of ~ 50 kD. From a combination of molecular sieving and ultracentrifugation studies, a calculated Mr of 61.4 kD is obtained for native mouse scatter factor, a value which agrees well with the Mr estimates obtained by SDS-PAGE (62-67 kD). Mouse fibroblast scatter factor is a heparin-binding, basic protein (pI 8.5-9.5) which contains N-linked carbohydrates which are not, however, essential for activity. The factor has no metallo- or serine protease activity and there is no evidence so far that its junctional-breaking activity involves proteolytic cleavage of surface molecules on target cells. Scatter factor is either identical or closely related to hepatocyte growth factor/hepatopoietin A (a potent mitogen for rat hepatocytes recently purified from human and rabbit serum and rat platelets). The factor is thus an effector of mesenchymal-epithelial interactions which affects the movement or the growth of different epithelia.


Hepatocyte Growth Factor MDCK Cell Scatter Factor Potent Mitogen Serine Protease Activity 
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  1. Adams, J. C., Furlong, R. A., and Watt, F. M. (1991) Production of scatter factor by ndk, a strain of epithelial cells, and inhibition of scatter factor activity by suramin. J. Cell Sci., 98: 385–394.Google Scholar
  2. Behrens, J., Weidner, K. M., Frixen, U. H., Schipper, J. H., Sachs, M., Arakaki, N., Daikuhara, Y., and Birchmeier, W. (1991) The role of E-cadherin and scatter factor in tumor invasion and cell motility, in: Cell Motility Factors, ed. I. D. Goldberg. Birkhäuser Verlag, Basel, pp. 109–126. (this volume)Google Scholar
  3. Bhargawa, M. M., Li, Y., Joseph, A., Pendergast, M., Hofmann, R., Rosen, E. M., and Goldberg, I. D., Purification, characterization and mechanism of action of scatter factor from human placenta, in: Cell Motility Factors, ed. I. D. Goldberg. Birkhäuser Verlag, Basel, pp. 63–75. (this volume)Google Scholar
  4. Coffer, A. I., Fellows, J., Young, S., Pappin, D., and Rahman, D. (1991) Purification and characterisation of biologically active scatter factor from ras-transformed NIH 3T3 conditioned medium.Google Scholar
  5. Dowrick, P. G., and Warn, R. M. (1991) The cellular response to factors which induce motility in mammalian cells, in: Cell Motihty Factors, ed. I. D. Goldberg. Birkhäuser Verlag, Basel, p. 89–108.Google Scholar
  6. Furlong, R. A., Biochem. J., in press. (1991) personal communication.Google Scholar
  7. Gherardi, E., and Stoker, M. (1990) Scatter factor and hepatocytes. Nature 346: 228.CrossRefGoogle Scholar
  8. Gherardi, E., Gray, J., Stoker, M., Perryman, M., and Furlong, R. (1989) Purification of scatter factor, a fibroblast-derived basic protein that modulates epithehal interactions and movement. Proc. natl. Acad. Sci. USA 86: 5844–5848.CrossRefGoogle Scholar
  9. Gohda, E., Tsubouchi, H., Nakayama, H., Hirono, S., Sakiyama, O., Takahashi, K., Miyazaki, H., Hashimoto, S., and Daikuhara, Y. (1988) Purification and partial characterization of hepatocyte growth factor from plasma of a patient with fulminant hepatic failure. J. Clin. Invest. 81: 414–419.CrossRefGoogle Scholar
  10. Higashio, K., Shima, N., Goto, M., Itagaki, Y., Nagao, M., Yasuda, M., and Morinaga, T. (1990) Identity of a tumour cytotoxic factor from human fibroblasts and hepatocyte growth factor. Biochem. Biophys. Res. Commun. 170: 397–404.CrossRefGoogle Scholar
  11. Miyazawa, K., Tsubouchi, H., Naka, D., Takahashi, K., Okigaki, M., Arakaki, N., Nakayama, H., Hirono, S., Sakiyama, O., Takahashi, K., Godha, E., Daikuhara, Y., and Kitamura, N. (1989) Molecular cloning and sequence analysis of cDNA for human hepatocyte growth factor. Biochem. biophys. Res. Commun. 163: 967–973.CrossRefGoogle Scholar
  12. Nakamura, T., Nawa, K., Ichihara, A., Kaise, N., and Nishino, T. (1987) Purification and subunit structure of hepatocyte growth factor from rat platelets. Fed. Eur. Biochem. Soc. Lett. 224: 311–316.CrossRefGoogle Scholar
  13. Nakamura, T., Nishizawa, T., Hagiya, M., Seki, T., Shimonishi, M., Sugimura, A., Tashiro, K., and Shimizu, S. (1989) Molecular cloning and expression of human hepatocyte growth factor. Nature 342: 440–443.CrossRefGoogle Scholar
  14. Oncley, J. L. (1941) Evidence from physical chemistry regarding the size and shape of protein molecules from ultra-centrifugation, diffusion, viscosity, dielectric dispersion, and double refraction of flow. Ann. N. Y. Acad. Sci. 41: 121–150.CrossRefGoogle Scholar
  15. Rosen, E. M., Grant, D., Kleinman, H., Jakens, S., Donovan, M. A., Setter, E., Luckett, P. M., and Carley, W. (1991) Scatter factor stimulates migration of vascular endothelium and capillary-like tube formation, in: Cell Motility Factor, ed. I. D. Goldberg. Birkhäuser Verlag, Basel, pp. 76–88. (this volume)Google Scholar
  16. Rosen, E. M., Goldberg, I. D., Kacinski, B. M., Buckholz, T., and Vinter, D. W. (1989) Smooth muscle releases an epithelial cell scatter factor which binds to heparin. In Vitro Cell. Dev. Biol. 25: 163–173.Google Scholar
  17. Rosen, E. M., Meromsky, L., Romero, R., Setter, E., and Golderg, I. (1990a). Human placenta contains an epithelial scatter protein. Biochem. biophys. Res. Commun. 168: 1082–1088.CrossRefGoogle Scholar
  18. Rosen, E. M., Meromsky, L., Setter, E., Vinter, D. W., and Goldberg, I. D. (1990b) Smooth-muscle-derived factor stimulates mobility of human tumour cells. Invas. Metastasis 10: 49–64.Google Scholar
  19. Rosen, E. M., Meromsky, L., Setter, E., Vinter, D. W., and Goldberg, I. D. (1990c) Purified scatter factor stimulates epithelial and vascular endothelial cell migration. Proc. Soc. exp. Biol. Med. 195: 34–43.Google Scholar
  20. Rosen, E. M., Meromsky, L., Setter, E., Vinter, D. W., and Goldberg, I. D. (1990d) Quantitation of cytokine-stimulated migration of endothelium and epithelium by a new assay using microcarrier beads. Exp. Cell Res. 186: 22–31.CrossRefGoogle Scholar
  21. Rubin, J. S., Chan, A. M.-L., Bottaro, D. P., Burgess, W. H., Taylor, W. G., Cech, A. C., Hirschfield, D. W., Wong, J., Miki, T., Finch, P. W., and Aaronson, S. A. (1991) A broad spectrum human lung fibroblast-derived mitogen is a variant of hepatocyte growth factor. Proc. natl. Acad. Sci., 88: 415–419.CrossRefGoogle Scholar
  22. Schor, S., Grey, A. M., Picardo, M., Schor, A. M., Howell, A., Ellis, I., and Rushton, G. (1991) Heterogeneity among fibroblasts in the production of migration stimulating factor (MSF): Implications for cancer pathogenesis, in: Cell Motihty Factors, ed. I. D. Goldberg. Birkhaüser Verlag, Basel, pp. 127–146. (this volume)Google Scholar
  23. Sporn, M. B., and Roberts, A. B. (1988) Peptide growth factors are multifunctional. Nature, 332: 217–219.CrossRefGoogle Scholar
  24. Stern, C., Ireland, G. W., Herrick, S. E., Gherardi, E., Gray, J., Perryman, M., and Stoker, M. (1990) Epithelial scatter factor and development of the chick embryonic axis. Development, in press.Google Scholar
  25. Stoker, M. (1984) Junctional competence in clones of mammary epithelial cells, and modulation by conditioned medium. J. cell. Physiol. 121: 174–183.CrossRefGoogle Scholar
  26. Stoker, M., and Gherardi, E. (1987) Factors affecting epithelial interactions, in: Junctional Complexes of Epothelial Cells, Ciba Foundation Symposium, Vol. 125. Wiley, Chichester, pp. 217–239.Google Scholar
  27. Stoker, M., and Gherardi, E. (1989) Scatter factor and other regulators of cell mobility. Br. med. Bull. 45: 481–491.Google Scholar
  28. Stoker, M., and Gherardi, E. (1991) Regulation of cell movement: the motogenic cytokines. Biochim. Biophys. Acta, 1072: 81–102.Google Scholar
  29. Stoker, M., Gherardi, E., Perryman, M., and Gray, J. (1987) Scatter factor is a fibroblast-derived modulator of epithelial cell mobility. Nature 327: 239–242.CrossRefGoogle Scholar
  30. Stoker, M., and Perryman, M. (1985) An epithehal scatter factor released by embryo fibroblasts. J. Cell Sci. 77: 209–223.Google Scholar
  31. Stracke, M. L., Aznavoorian, S. A., Beckner, M. E., Liotta, L. A., and Schiffmann, E. (1991) Cell motility, a principal requirement for metastasis, in: Cell Motility Factors, ed. I. D. Goldberg. Birkhauser Verlag, Basel, pp. 147–162. (this volume)Google Scholar
  32. Weidner, M. K., Behrens, J., Vandekerckhove, J., and Birchmeyer, W. (1990) Scatter factor: Molecular characteristics and effect on the invasiveness of epithehal cells. J. Cell Biol. 111: 2097–2108.CrossRefGoogle Scholar
  33. Zarnegar, R., and Michalopoulos, G. (1989) Purification and biological characterization of human Hepatopoietin A, a polypeptide growth factor for hepatocytes. Cancer Res. 49: 3314–3320.Google Scholar

Copyright information

© Birkhäuser Verlag Basel/Switzerland 1991

Authors and Affiliations

  • Ermanno Gherardi
    • 1
    • 2
  • Arnold Coffer
    • 1
    • 3
  1. 1.Imperial Cancer Research Fund LaboratoriesUK
  2. 2.Department of Medicine, Addenbrooke’s HospitalCambridge UniversityCambridgeUK
  3. 3.Protein Isolation and Cloning LaboratoryLincoln’s Inn FieldLondonEngland

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