Hepatocyte Growth Factor/Scatter Factor Binds with High Affinity to Dermatan Sulfate*
- Malcolm Lyon‡§,
- Jon A. Deakin‡,
- Hassan Rahmoune‖,
- David G. Fernig‖,
- Toshikazu Nakamura** and
- John T. Gallagher‡
- From the ‡Cancer Research Campaign & University of Manchester, Department of Medical Oncology, Christie Hospital National Health Service Trust, Manchester M20 4BX, United Kingdom, the‖School of Biological Sciences, University of Liverpool, Liverpool L69 7ZB, United Kingdom, and the **Division of Biochemistry, Biomedical Research Center, Osaka University Medical School, Osaka 565, Japan
Abstract
We have demonstrated by affinity chromatography that hepatocyte growth factor/scatter factor (HGF/SF) binds strongly to dermatan sulfate (DS), with a similar ionic strength dependence to that previously seen with heparan sulfate (HS). Analysis of binding kinetics on a biosensor yields an equilibrium dissociation constant,K D, of 19.7 nm. This corresponds to a 10–100-fold weaker interaction than that with HS, primarily due to a faster dissociation rate of the complex. The smallest DS oligosaccharide with significant affinity for HGF/SF by affinity chromatography appears to be an octasaccharide. A sequence comprising unsulfated iduronate residues in combination with 4-O-sulfated N-acetylgalactosamine is sufficient for high affinity binding. The presence of 2-O-sulfation on the iduronate residues does not appear to be inhibitory. These observations concur with our previous suggestions, from analyses of HS binding (Lyon, M., Deakin, J. A., Mizuno, K., Nakamura, T., and Gallagher, J.T. (1994) J. Biol. Chem. 269, 11216–11223), that N-sulfation of hexosamines and 2-O-sulfation of iduronates are not absolute requirements for glycosaminoglycan binding to HGF/SF. This is the first described example of a high affinity interaction between a growth factor and DS, and is likely to have significant implications for the biological activity of this paracrine-acting factor.
Footnotes
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↵* This work was supported by grants from the Cancer Research Campaign (to M. L., J. A. D., and J. T. G.), the North West Cancer Research Fund (to H. R. and D. G. F.), and the Mizutani Foundation for Glycoscience.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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↵§ To whom correspondence should be addressed: Dept. of Medical Oncology, Christie CRC Research Centre, Christie Hospital NHS Trust, Wilmslow Rd., Manchester M20 4BX, United Kingdom. Tel.: 44-161-446-3202; Fax: 44-161-446-3269; E-mail:MLyon{at}picr.man.ac.uk.
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↵1 The abbreviations used are: HGF/SF, hepatocyte growth factor/scatter factor; GAG, glycosaminoglycan; HS, heparan sulfate; HSPG, heparan sulfate proteoglycan; DS, dermatan sulfate; DSPG, dermatan sulfate proteoglycan; CS, chondroitin sulfate; FGF, fibroblast growth factor; HPLC, high performance liquid chromatography; PBS, phosphate-buffered saline; dp, degree of polymerization (i.e. number of monosaccharides); GlcA, β-d-glucuronate; IdoA, α-l-iduronate; HexA, unspecified hexuronate; ΔHexA, Δ4,5-unsaturated hexuronate; IdoA(2-OSO3), α-l-iduronate 2-sulfate; GalNAc, β-d-N-acetylgalactosamine; GalNAc(4-OSO3), β-d-N-acetylgalactosamine 4-sulfate; GalNAc(6-OSO3), β-d-N-acetylgalactosamine 6-sulfate; GalNAc(4,6-OSO3), β-d-N-acetylgalactosamine 4,6-disulfate; MDCK, Madin-Darby canine kidney.
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↵2 H. Rahmoune, P. S. Rudland, J. T. Gallagher, and D. G. Fernig, submitted for publication.
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↵3 Rahmoune, H., Chen, H.-L., Gallagher, J. T., Rudland, P. S., and Fernig, D. G. (1998) J. Biol. Chem., in press.
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- Received July 28, 1997.
- Revision received October 9, 1997.
- The American Society for Biochemistry and Molecular Biology, Inc.
