Lyon M, Deakin J A, Rahmoune H, Fernig D G, Nakamura T, Gallagher J T
Cancer Research Campaign & University of Manchester, Department of Medical Oncology, Christie Hospital National Health Service Trust, Manchester M20 4BX, United Kingdom.
J Biol Chem. 1998 Jan 2;273(1):271-8. doi: 10.1074/jbc.273.1.271.
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, KD, 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.
我们通过亲和层析证明，肝细胞生长因子/分散因子（HGF/SF）与硫酸皮肤素（DS）强烈结合，其离子强度依赖性与先前观察到的硫酸乙酰肝素（HS）相似。在生物传感器上分析结合动力学得出平衡解离常数KD为19.7 nM。这对应于比与HS的相互作用弱10 - 100倍的相互作用，主要是由于复合物的解离速率更快。通过亲和层析对HGF/SF具有显著亲和力的最小DS寡糖似乎是八糖。由未硫酸化的艾杜糖醛酸残基与4 - O - 硫酸化的N - 乙酰半乳糖胺组成的序列足以实现高亲和力结合。艾杜糖醛酸残基上2 - O - 硫酸化的存在似乎没有抑制作用。这些观察结果与我们之前从HS结合分析中得出的建议一致（Lyon, M., Deakin, J. A., Mizuno, K., Nakamura, T., and Gallagher, J.T. (1994) J. Biol. Chem. 269, 11216 - 11223），即己糖胺的N - 硫酸化和艾杜糖醛酸的2 - O - 硫酸化不是糖胺聚糖与HGF/SF结合的绝对要求。这是首次描述的生长因子与DS之间高亲和力相互作用的例子，并且可能对这种旁分泌作用因子的生物活性具有重要意义。
