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標題: | 尋找與人類甲基轉移酶SMYD交互作用之可能蛋白 Investigation of Protein-Protein Interaction of SET and MYND Domain-containing proteins |
作者: | 薛翔尹 Hsiang-Yin Hsueh |
指導教授: | 鄧述諄 Shu-Chun Teng |
關鍵字: | SMYD protein,非組蛋白甲基化,賴氨酸甲基轉移?,LDH,CCT2,IGFBP4, SMYD family,Non-histone protein methylation,lysine methyltransferase,LDH,CCT2,IGFBP4, |
出版年 : | 2018 |
學位: | 碩士 |
摘要: | 賴氨酸甲基轉移酶(KMT)的SMYD蛋白家族由五個成員組成,每個成員皆具有SET結構域和MYND結構域。它們最早被視為一般組蛋白甲基轉移酶,在不同的細胞過程中像是染色絲重塑和基因表達發揮重要作用。近來愈來愈多研究指出特定癌症生長及心血管疾病與SMYD的過表現或是失能相關。因此,為了了解SMYD家族在細胞中未知的功能,我們利用質譜分析、螢光顯微鏡、酵母菌雙雜合系統等技術尋找SMYD交互作用蛋白,藉此發掘SMYD的新功能。
結果顯示SMYD1與乳酸脫氫酶-LDHA和LDHB具有交互作用。在SMYD1剔除的RD細胞中發現LDHA和LDHB蛋白質量的下降,並且在體外活性測定中觀察到LDH的活性增加,顯示SMYD1可能參與在無氧呼吸中。 其次,T蛋白複合物的亞基之一CCT2特異性影響SMYD5的表現量,揭露SMYD5新的蛋白質摺疊路徑。綜合上述,這項研究指出SMYD可能參與其中的新路徑,有助於了解SMYD在細胞中的生理功能。 The SMYD (SET and MYND domain containing proteins) family of lysine methyltransferases (KMTs) is composed of five members. They were first identified as general histone methyltransferase, played essential roles in different cellular processes including chromatin remodeling and gene expression. Recently, more and more research indicated that SMYD overexpression or dysfunction is associated with cancer development and several cardiac diseases. To gain more knowledge of this relatively uncharacterized family, we turn to Mass spectrometry, Fluorescence microscope and Yeast-two hybrid system to find potential interacting proteins of SMYD and further discover their new function in cells. Firstly, we found that SMYD1 has interaction with the subunit of lactate dehydrogenase-LDHA and LDHB. Moreover, in SMYD1 knockdown RD cell line, the protein level of both LDHA and LDHB decreased and the activity of the LDH increased in the in vitro activity assay, indicates that SMYD1 may involve in aerobic respiration regulation. Secondly, the subunit of the T protein complex CCT2, affected the protein level of SMYD5 specifically. The result reveals a new protein folding pathway of SMYD5. Thirdly, we observed that in SMYD4 knockdown MCF7 cell line the phosphorylation of IGF1R decreased and in SMYD4 knockdown SHSY5Y cell line the phosphorylation level of Akt decreased, which showed that SMYD4 may play a role in the insulin like growth factor signaling pathway. Altogether, this study shed light on several pathways that SMYD may participate in. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/77599 |
DOI: | 10.6342/NTU201801674 |
全文授權: | 未授權 |
顯示於系所單位: | 微生物學科所 |
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