| Summary: | 碩士 === 國立清華大學 === 化學系 === 96 === The fibroblast growth factors (FGFs) are members of a large family of signaling molecules with many diverse biological activities that include cellular growth, proliferation and tumor growth. FGF-2 is a single-chain heparin-binding polypeptide and has been detected in normal and malignant cells and shown for its mitogenic and angiogenic activity with a consequent crucial role in cell differentiation and development. In particular, FGF-2 can be secreted to the extracellular medium where it switches on signal transduction pathways by binding to and activation of cell-surface receptors.
Angiogenic inhibitors recently have been demonstrated as potential anticancer agents. Moreover, inhibition FGFs angiogenic activity has been suggested as a crucial target for the development of anticancer agents. In this study, we describe an organic compound -SSR1281299E (abbreviate SSR) that binds to FGF-2 and inhibits the interaction between FGF-2 and its receptor, which is related to anti-angiogenesis and antitumor activity.
The binding region of SSR on the FGF-2 protein was characterized using 1H-15N HSQC perturbation experiments. Biophysical experiment of fluorescence and far UV CD (circular dichroism) suggested that SSR was able to stabilize hFGF-2. To determine the conformation of FGF-2 in the bound form, a variety of 3D NMR experiments include HNCA, HNCO, CBCA(CO)NH, HBHA(CO)NH and HCCH-TOCSY were used to accomplish the 1H,15N and 13C assignments in the bound form of FGF-2. After integrating all constraints that include 15N/13C- edited NOESY, dihedral angle, chemical shift index (CSI) and hydrogen bond, the 3D solution structure of hFGF-2 in complex with SSR was solved by ARIA-CNS. Moreover, To understand the molecular level binding site between FGF-2 and SSR, 13C＆15N-filtered-13C edited NOESY technique were applied to obtain the information of intermolecular NOE from complex system. By means of molecular modeling procedures, the structure-activity relationships between FGF-2 and SSR have been analyzed on the basis of the HADDOCK generated structures.
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