人類S100蛋白質家族是由一群小分子的酸性蛋白所組成，這個蛋白質家族在結構上具有高度相似性，它們皆具有兩個EF-hand motif能與鈣離子結合，在與鈣離子結合後會改變其構形並與目標蛋白產生交互作用，進而引發一連串的生化反應，S100A4為S100蛋白質家族其中一員且EGF蛋白是能與S100A4結合的標的蛋白之一。人類表皮生長因子（Human Epidermal Growth Factor, hEGF）可促使多種生物細胞（包括腫瘤細胞）的增生分化，而此生物活性是透過EGF蛋白與細胞膜表面上的特定受體（EGFR）結合進而誘導產生一系列訊息傳導所致。在先前的研究中，作者利用西方點墨法指出S100A4蛋白會與EGF蛋白產生交互作用。氨來呫諾（Amlexanox, AMX）是一種特殊的抗發炎及抗過敏的藥物，目前主要是用來治療口腔潰瘍，在本實驗中我們以HSQC滴定證實AMX與S100A4之間有交互作用。我們利用了螢光光譜儀（fluorescence spectroscopy）及核磁共振光譜儀（NMR spectroscopy）證明了AMX與EGF兩者分別與S100A4之間的交互作用關係，再加上我們解出的複合體水溶液結構，接著以WST-1 assay研究三者之間在生物活性方面的調控，綜合以上研究成果希望能進一步的探討S100A4、EGF與AMX三者在生物活性調控機轉之間的關係。

Human S100 proteins belong to a family of small, acidic protein which shares high structure similarities with each other: they all have two EF-hand motifs and could bind calcium. When S100A4 binds with calcium, it will change its conformation and interact with their target protein. Human epidermal growth factor ( hEGF ) is the target protein which is also one of the high affinity ligands of EGFR. EGF/EGFR system promotes cell survival, growth and differentiation via the activation of several integrated signaling pathways. In recent studies, scientists used western blotting to show that S100A4 could interact with EGF. Amlexanox is an anti-inflammatory and antiallergic drug used to treat recurrent aphthous ulcers. In this study, we found AMX interact with S100A4 using HSQC titrations. We elucidated the interactions of S100A4 with EGF and AMX using fluorescence spectroscopy and NMR spectroscopy. We solved the solution complex structures and investigated the bioactivities using WST-1 assay. The results we reported could help to investigate the biological mechanism among S100A4, EGF and AMX.

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