title: Interaction of hepatocyte growth factor with heparan sulfate. Elucidation of the major heparan sulfate structural determinants.
abstract: We have demonstrated by affinity chromatography that hepatocyte growth factor (HGF) binds strongly to heparan sulfate (HS). This substantiates previous suggestions that cell-surface heparan sulfate proteoglycans constitute the so-called low affinity cellular binding sites for HGF. Using a recombinant human HGF affinity column, we have analyzed the effects of various specific chemical and enzymatic modifications/depolymerizations of HS on its affinity in order to elucidate the polysaccharide structural determinants. Interaction is shown to be only slightly affected by digestion with heparinase I or III or by replacement of N-sulfates with N-acetyl groups. This suggests a specific role for sulfated domains containing nonsulfated IdceA residues, with only a small contribution from N-sulfates and IdceA(2-OSO3) residues. In addition, disaccharide analyses of various HGF-binding oligosaccharides indicate that affinity is more closely associated with 6-O-sulfation of GlcNSO3 residues than with sulfation at any other position. Although interaction can be demonstrated with heparinase III-resistant oligosaccharides as small as hexasaccharides, the highest affinity was found with oligosaccharides containing a minimum of 10-12 monosaccharides. The structural specificity of the HGF-HS interaction is thus shown to be radically different from that previously described for the basic fibroblast growth factor-HS interaction.
authors.affiliation: Cancer Research Campaign, Christie Hospital, Manchester, United Kingdom.
authors.forename: M
authors.initials: M
authors.lastname: Lyon
authors.forename: J A
authors.initials: JA
authors.lastname: Deakin
authors.forename: K
authors.initials: K
authors.lastname: Mizuno
authors.forename: T
authors.initials: T
authors.lastname: Nakamura
authors.forename: J T
authors.initials: JT
authors.lastname: Gallagher
mesh_terms: D000818:Animals; D002240:Carbohydrate Sequence; D002846:Chromatography, Affinity; D004187:Disaccharides / Q000737:chemistry / Q000302:isolation & purification; D005347:Fibroblasts / Q000378:metabolism; D019812:Heparan Sulfate Proteoglycans; D006493:Heparin / Q000378:metabolism; D006497:Heparan Sulfate / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D017228:Hepatocyte Growth Factor / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D008099:Liver / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D011509:Proteoglycans / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D004187:Disaccharides; D019812:Heparan Sulfate Proteoglycans; D006493:Heparin; D006497:Heparan Sulfate; D017228:Hepatocyte Growth Factor; D009844:Oligosaccharides; D011509:Proteoglycans; D011994:Recombinant Proteins