Surface plasmon resonance analysis to evaluate the importance of heparin sulfate groups' binding with human aFGF and bFGF |
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| Authors: | Email author" target="_blank">Wu?Xiao-fengEmail author Xu?Ya-xiang Shen?Guo-xin Kamei?Kaeko Takano?Ryo Hara?Saburo |
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| Institution: | (1) College of Animal Sciences, Zhejiang University, Huajiachi Campus, 310029 Hangzhou, China;(2) Department of Applied Biology, Kyoto Institute of Technology, 606 Kyoto, Japan |
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| Abstract: | 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 mixtures 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 be used for analysing the effect of sulfate groups in heparin on the binding with other human
FGF members or other heparin-binding proteins.
The project supported by the Scientific Research Foundation for Returned Overseas Chinese Scholars, State Education Ministry
of China and Zhejiang Provincial Natural Science Foundation of China (301306) |
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| Keywords: | Surface plasmon resonance analyses Heparin Sulfate groups Human aFGF and bFGF |
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