Kishimoto Satoko, Nakamura Shingo, Hattori Hidemi, Nakamura Shin-Ichiro, Oonuma Fumie, Kanatani Yasuhiro, Tanaka Yoshihiro, Mori Yasutaka, Harada Yasuji, Tagawa Masahiro, Ishihara Masayuki
Research Institute, National Defense Medical College, 3-2 Namiki, Tokorozawa, Saitama, Japan.
J Biochem. 2009 Mar;145(3):275-8. doi: 10.1093/jb/mvn169. Epub 2008 Dec 12.
Binding affinities of chemically modified heparins for human stem cell factor (SCF) were examined using fragmin/protamine microparticles (F/P MPs) and an enzyme-linked immunosorbent assay (ELISA). The binding of SCF to F/P MP-coated plates was inhibited with high concentrations of heparin and fragmin, but not others. The binding of SCF was also inhibited with 0.55 M or higher concentrations of NaCl in the medium. These results suggested that a high content of all three sulfate groups in repeating disaccharide units is required for interaction with SCF. Furthermore, pre-immobilized SCF on F/P MP-coated plates significantly stimulated proliferation of a human erythroleukemia cell line.
使用抑肽酶/鱼精蛋白微粒(F/P MPs)和酶联免疫吸附测定(ELISA)检测化学修饰肝素对人干细胞因子(SCF)的结合亲和力。高浓度的肝素和抑肽酶可抑制SCF与F/P MP包被板的结合,而其他物质则无此作用。培养基中0.55 M或更高浓度的NaCl也可抑制SCF的结合。这些结果表明,重复二糖单元中所有三个硫酸基团的高含量是与SCF相互作用所必需的。此外,预先固定在F/P MP包被板上的SCF可显著刺激人红白血病细胞系的增殖。