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請使用永久網址來引用或連結此文件:
http://libir.tmu.edu.tw/handle/987654321/4226
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| 題名: | 新穎蛋白激脢 mSBK 功能的鑑定
Characterization of a novel brain predominant protein kinase, mouse SH3 binding kinase (mSBK) |
| 作者: | 黃一斌
Yi-Pin Huang |
| 貢獻者: | 醫學科學研究所 |
| 關鍵詞: | 絲胺酸
蘇胺酸激酶
mSBK
MYH9
serine/threonine |
| 日期: | 2009 |
| 上傳時間: | 2009-08-27 17:27:02 (UTC+8) |
| 摘要: | 蛋白激酶在調節大腦功能上扮演著重要角色，其中包括：神經細胞的分化 (neuronal differential)、神經的多變性 (neuronal plasticity)、LTP (long-term potentiation)、LTD (long-term depression) 和神經傳導物質 (neurotransmitter release) 的釋放等。實驗室過去在斑馬魚的研究中發現一個新穎的 serine /threonine 蛋白激酶 BSK146，結果顯示此蛋白激酶可能參與後腦發育；而此新穎的蛋白激酶依胺基酸序列相似性分析發現，在大鼠研究中也有一序列相似的蛋白激酶 sbk，過去的研究報告指出此蛋白激酶 C 端有一 proline-rich 的區域。為了要鑑定新穎蛋白激酶的功能，我們以老鼠腦部的 cDNA 選殖出小鼠的 sbk 基因 (msbk)，來探討這一新穎蛋白激酶可能的機制。利用 pull-down 的實驗，在星狀細胞細胞萃取液中發現 MYH9 (non-muscle myosin heavy chain IIA) 會與 mSBK 結合；此外，為了要瞭解 msbk 基因的調控情形，我們分析了 msbk 基因上游 5kb 的片段，發現許多具有神經組織專一性結合的轉錄因子結合位置。在 promoter 活性分析的實驗中發現，在神經細胞內 mSBK 片段七的區域具有 promoter 活性。而免疫染色證實在神經細胞當中存在著 MYH9 蛋白。在本研究中，我們發現了會與 mSBK 結合的蛋白以及預測會與轉錄因子結合的位置。是否 mSBK 透過與 MYH9 結合進而影響下游蛋白以調控腦部功能或是在發育階段使神經細胞遷移到適當位置值得未來進一步的探討。
Protein kinases play important roles in the regulation of several brain-specific functions, including neuronal differential, neuronal plasticity, LTP (long-term potentiation)，LTD (long-term depression) and neurotransmitter release. In the previously studies, we have identified a novel serine/threonine protein kinase, BSK146, in the brain of zebrafish. In the present data suggest that BSK146 may involve in hindbrain development. By using the BLAST program to search the currently available database, we found a BSK146 homologue, rat SH3-binding kinase (sbk), with distinct kinase domain and proline-rich C-terminal region. To identify this novel protein kinase function, we isolated and characterized a mice protein kinase msbk, which is homologous to that of rat SH3-binding kinase (sbk), and explore this novel protein kinase possible mechanism. Using pull-down approach, we have identified a candidate msbk interaction protein in astrocytes cell lysate, MYH9 (non-muscle myosin heavy chain IIA). In addition to elucidate the regulation of msbk gene expression, we analyzed the 5kb fragment corresponding to the 5’-upstream region of msbk gene, revealed numerous putative binding sites for transcription factors. Promoter analysis proved that the mSBK 7th fragment had promoter activity in nerve cells. By using the immuno-fluorescence analysis also found the MYH9 in nerve cells. In this study, we have identified a candidate interaction molecule for msbk and revealed numerous putative regulation elements. Whether the mSBK were associated with MYH9 to affect the downstream protein and regulated the other specific brain functions or nerve cells to migrate properly during developmental stages need to be further explored. |
| 資料類型: | thesis |
| 顯示於類別: | [醫學科學研究所] 博碩士論文
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| tmu-98-M120096009-1.pdf | | 2182Kb | Adobe PDF | 562 | 檢視/開啟 |
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