Characterization of the surface immobilized synthetic heparin binding domain derived from human fibroblast growth factor-2 and its effect on osteoblast differentiation
Characterization of the surface immobilized synthetic heparin binding domain derived from human fibroblast growth factor-2 and its effect on osteoblast differentiation
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DOI：
10.1002/jbm.a.31351
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发表时间：
2007-12-15
影响因子：
4.9
通讯作者：
Park, Yoon-Jeong
中科院分区：
文献类型：
--
作者：
Lee, Jue-Yeon;Choo, Jung-Eun;Park, Yoon-Jeong
成纤维细胞生长因子（FGF）-2通过在肝素蛋白聚糖存在下与细胞表面FGF受体（FGFR）结合来调节多种细胞功能，例如增殖和分化。FGF-2是已知的肝素结合生长因子，但肝素结合位点的定位至今尚未完全研究。我们使用FGF-2的两个潜在肝素结合结构域，残基105-111（17105，YKRSRYT）和119135（17119，KRTGQYKLGSKTGPGQK）。多肽可以稳定地固定在组织培养板表面。使用固相结合试验，我们证明了这两种肽对肝素具有更高的结合亲和力相比，非结合对照序列。这些网站的生物学意义进行了测试的细胞附着和成骨细胞分化的研究。细胞对肽F105和F119的附着以剂量依赖性方式增加。肝素和肝素酶处理降低了细胞对F105和F119的粘附。这表明F105和F119都与细胞表面硫酸乙酰肝素蛋白聚糖相互作用，表明FGF-2具有两个肝素结合位点。此外，成骨细胞的分化，证实了ALP 13的活性和矿化，增加了表面固定肽F105和F119。总之，这些肝素结合肽可以作为生物制剂，增强成骨细胞分化以及表面修饰工具，在组织再生领域，特别是骨再生。(c)2007 Wiley Periodicals，Inc.
Fibroblast growth factor (FGF)-2 regulates a variety of cellular functions, such as proliferation and differentiation, by binding to cell surface FGF receptors (FGFRs) in the presence of heparin proteoglycans. FGF-2 is known as a heparin-binding growth factor, but the localization of the heparin binding site has not been fully investigated until now. We used two potential heparin binding domains of FGF-2, the residues 105-111 (17105, YKRSRYT) and 119135 (17119, KRTGQYKLGSKTGPGQK). Peptides could be stably immobilized onto the surface of tissue culture plates. Using solid phase binding assays, we demonstrated that both peptides had higher binding affinity toward heparin compared with nonbinding control sequence. The biological significance of these sites was tested by cell attachment and osteoblast differentiation studies. Cell attachment to the peptides F105 and F119 increased in a dose-dependent manner. Heparin and heparinase treatments decreased cell adhesion to both F105 and F119. This demonstrates that both F105 and F119 interact with cell-surface heparan sulfate proteoglycans, suggesting that FGF-2 has two heparin binding sites. In addition, osteoblast differentiation, confirmed by ALPase activity and mineralization, was increased by surface immobilized peptide F105 and F119. Taken together, these heparin binding peptides could be applied as biological agents enhancing osteoblast differentiation as well as surface modification tools in the tissue regeneration area, especially for bone regeneration. (c) 2007 Wiley Periodicals, Inc.