https://scholars.lib.ntu.edu.tw/handle/123456789/160449
標題: | 鑑定蛋白酪胺酸磷酸?(非受體二十二型)之細胞核定位訊號 Identification of Nuclear Localization Signal (NLS) in PTPN22 |
作者: | 孫宜緯 Sun, Yi-Wei |
關鍵字: | 蛋白酪胺酸磷酸?非受體二十二型;PTPN22;NLS | 公開日期: | 2010 | 摘要: | 蛋白酪胺酸磷酸酶非受體二十二型為在細胞質中負向調控T細胞訊息路徑的酪胺酸磷酸酶。我們實驗室近來在細胞核中發現此酵素。然而,蛋白酪胺酸磷酸酶非受體二十二型進入細胞核的機制仍未明。先前已知帶有細胞核定位訊號的蛋白質可以被運送進細胞核中。我們證實蛋白酪胺酸磷酸脢非受體二十二型的區域C為潛在的細胞核定位訊號,而且區域C對於細胞核運送是必須存在的。帶有區域C被刪除的蛋白酪胺酸磷酸脢非受體二十二型之螢光表現質體在轉染細胞後,發現其螢光無法分布於細胞核中。從點突變實驗中,我們也證實蛋白酪胺酸磷酸脢非受體二十二型的A和B點突變之後,將會損壞此蛋白質進入細胞核的能力。進一步研究負責運送蛋白酪胺酸磷酸脢非受體二十二型進入細胞質的胺基酸序列之實驗中,我們將區域I或/和區域II和雙倍的綠色螢光蛋白克隆在一起,製造出融合蛋白,驗證這些區域對於將綠色螢光蛋白送運進入細胞核的能力為何。從實驗結果發現,蛋白酪胺酸磷酸酶非受體二十二型的區域I能將綠色螢光蛋白運送進入細胞核。我們的實驗指出蛋白酪胺酸磷酸酶非受體二十二型的區域I為細胞核定位訊號且在運送蛋白質進入細胞核中扮演十分重要的角色。 PTPN22 is a protein tyrosine phosphatase that participates in the negative regulation of T cell signaling in cytoplasm. Recently, our lab shows that PTPN22 could locate inside the nucleus. However, the molecular mechanism of PTPN22 in nuclear import is poorly understood. Proteins that import from the cytosol to the nucleus contain basic amino acids-rich nucleus localization signal (NLS). We demonstrated Region C of PTPN22 was a potential NLS and necessary for nuclear import. Cells transfected with fluorescent vectors encoding Region C-deleted PTPN22 showed clear nuclear distribution. Mutation analysis showed A and B mutations of PTPN22 are critical sites for nuclear import of PTPN22, because the Mutant A and B impaired the capability of nuclear transport of PTPN22. To further identify sequences responsible for nuclear import of PTPN22, Region I or/and Region II was fused with double GFP and examined their abilities to target GFP into the nucleus. The results showed that Region I of PTPN22 could import GFP into the nucleus. Our data indicated that the Region I of PTPN22 served as NLS and played a vital role in nuclear import. |
URI: | http://ntur.lib.ntu.edu.tw//handle/246246/248009 |
顯示於: | 免疫學研究所 |
檔案 | 描述 | 大小 | 格式 | |
---|---|---|---|---|
ntu-99-R97449010-1.pdf | 23.32 kB | Adobe PDF | 檢視/開啟 |
在 IR 系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。