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首页> 外文期刊>Journal of Zhejiang University Science: An international applied physics & engineering journal >Surface plasmon resonance analysis to evaluate the importance of heparin sulfate groups' binding with human aFGF and bFGF
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Surface plasmon resonance analysis to evaluate the importance of heparin sulfate groups' binding with human aFGF and bFGF

机译：表面等离子体共振分析，以评估硫酸肝素基团与人aFGF和bFGF结合的重要性

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摘要

Human acidic and basic fibroblast growth factors (aFGF and bFGF) are classic and well characterized members of the heparin-binding growth factor family. Heparin is generally thought to play an extremely important role in regulating aFGF and bFGF bioactivities through its strong binding with them. In order to unravel the mechanism of the interactions between heparin and FGFs, and evaluate the importance of heparin sulfate groups' binding with FGFs, surface plasmon resonance analyses were performed using IAsys Cuvettes System. Heparin and its regioselectively desulfated derivatives were immobilized on the cuvettes. aFGF and bFGF solutions with different concentrations were pipetted into the cuvettes and the progress of the interaction was monitored in real-time by Windows-based software, yielding kinetic and equilibrium constants for these interactions. In addition, in order to reduce the delicate difference among the cuvettes, inhibition analyses of mixture of FGFs and immobilized native heparin by modified heparins were also done. The data from these two methods were similar, indicating that all sulfate groups at 2-O, 6-O and N- in heparin were required for the binding to aFGF; and that their contribution to the binding was in the order 2-O, N- and 6-O-sulfate group. In contrast, definite contribution of the 6-O-sulfate group to the binding with bFGF was most apparent, while the other two sulfate groups appeared to be necessary in the order 2-O and N-sulfate group. These methods established here can he used for analysing the effect of sulfate groups in heparin on the binding with other human FGF members or other heparin-binding proteins.
机译：人酸性和碱性成纤维细胞生长因子（aFGF和bFGF）是肝素结合生长因子家族的经典且特征明确的成员。通常认为，肝素通过与aFGF和bFGF的强结合，在调节aFGF和bFGF的生物活性中起着极其重要的作用。为了阐明肝素与FGF之间的相互作用机制，并评估硫酸肝素基团与FGF结合的重要性，使用IAsys Cuvettes系统进行了表面等离子体共振分析。肝素及其区域选择性脱硫的衍生物被固定在比色皿上。将不同浓度的aFGF和bFGF溶液移入比色杯中，并通过基于Windows的软件实时监控相互作用的进程，并为这些相互作用产生动力学常数和平衡常数。此外，为了减少比色皿之间的细微差别，还进行了改性肝素对FGF和固定化天然肝素混合物的抑制分析。这两种方法的数据相似，表明肝素中2-O，6-O和N-处的所有硫酸盐基团都需要与aFGF结合。并且它们对结合的贡献为2-O，N-和6-O-硫酸根基团。相反，最明显的是6-O-硫酸根基团对与bFGF结合的明确贡献，而其他两个硫酸根基团似乎是必需的2-O和N-硫酸根基团。此处建立的这些方法可用于分析肝素中硫酸盐基团对与其他人FGF成员或其他肝素结合蛋白结合的影响。

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