Purification and crystallization of GST-MyD88

• Main Authors: Shih-De Yang, 楊士德
• Other Authors: 蔡麗珠
• Format: Others
• Language: zh-TW
• Published: 2008
• Online Access: http://ndltd.ncl.edu.tw/handle/s74356

碩士 === 國立臺北科技大學 === 有機高分子研究所 === 96 === Functional analysis of mammalian Toll-like receptors (TLRs) has revealed that they recognize specific patterns of microbial components that are conserved among pathogens. In signaling pathways via TLRs, a common adaptor, MyD88 (Myeloid differentiation primary-response protein 88), was first characterized as an essential component for the activation of innate immunity by all the TLRs. MyD88 is composed of a C-terminal TIR Domain and N-terminal death domain, which functions as an adaptor linking TLRs/IL-1Rs with downstream signaling molecules which is induced activation of NF-κB(nuclear factor-kappa B). In order to understand the protein-protein interaction and the function of MyD88, we plan to resolve the crystallographic structure of MyD88. Here, we describe how we purify and crystallize GST-MyD88. MyD88 was constructed into pGEX4T-1 vector and transformed into E. coli. BL21. The expression proteins were purified by a glutathione-S-transferase (GST)-affinity chromatography, and anion-exchange Q sepharose. Crystallization of the purified GST-MyD88 protein is in progress.