TLR/IL-1R途徑在對抗病原侵入與發炎反應裡扮演重要角色，此路徑的異常調控造成人類疾病。其中白細胞介素受體相關激酶 (IRAK) 家族在TLR/IL-1途徑中，扮演一個關鍵的調節者。在家族成員中，IRAKM作為負向調控的成員，其透過死亡結構域 (death domain) 之間作用形成一個巨大的複合物 (IRAKM-myddosome)，去執行訊息傳遞以及抑制訊息的調控。研究發現，IRAKM失控與哮喘 (asthma) 和慢性發炎(chronic inflammation) 相關。因此，為了瞭解參與在複合體中分子的交互作用，並對藥物開發提供新的見解。所以在我們的研究中，IRAKM-myddosome的重組蛋白成功地被建立且利用大腸桿菌系統進行目標蛋白表達，更進一步，利用親和性層析法和粒徑篩層析法，也成功得到純的蛋白質複合體。此外，也利用X光結晶學和小角度X光散射的方法去分析其蛋白質複合體。另一方面，利用穿透式電子顯微鏡觀察到IRAK-myddosome形成直徑約15 nm的顆粒。除此之外，在分子交互作用的聚合面上進行關鍵的點突變，我們得到許多蛋白質如何交互作用的資訊。總體而言，我們希望可以清楚地去瞭解，複合物中的蛋白質，彼此分子之間互相作用的機制，將有助於清楚了解NF-κB 訊息傳遞蛋白調控機制。

The TLR/IL-1R pathway plays an important role in anti-pathogen, inflammatory responses and their malfunction usually cause human diseases. The interleukin receptor-associated kinases (IRAKs) family act as key regulators in the TLR/IL-1R pathway. One of them IRAKM plays a negative regulatory role through death domain interaction to form the large complex called IRAKM-myddosome for triggering and downregulating the signaling. It has been reported the IRAKM dysregulation causes asthma, chronic inflammation. Hence, to understand the interaction pattern between these molecules may provide new insight for therapeutic agents. In my study, the recombinant protein complexes were cloned and co-expression by the Escherichia coli expression system, successfully. The two protein complexes were further purified by affinity and size exclusion chromatography. Besides, X-ray crystallography and Small Angle X-ray Scattering were applied to analyze the pure protein complexes. On the other hand, the transmission electron microscopy was used to observed IRAKM-myddosome formation particles with a diameter of about 15 nm. Besides, we get more information from the key vital residues in interface mutagenesis. Finally, we hope to understand clearly protein-protein interaction mechanism of the two complexes above in NF-κB signaling pathway.

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