PRMT1最早是藉由與哺乳類TIS21早期立即蛋白交互作用而被選殖出來，為哺乳類動物細胞中最主要的精胺酸甲基轉移酶，其提供了大於90%以上的精胺酸甲基化活性。為了了解PRMT1的酵素作用能力，我們以GST融合的PRMT1針對hnRNP A2此甲基化受質，在不同條件下進行甲基化反應，並以SDS-PAGE及fluorography分析。我們發現，二價陽離子的存在會影響PRMT1酵素催化能力，其中又以鈣離子有最顯著的影響，而單價陽離子則影響較小。
藉由與TIS21/BTG1 基因產物或是interferon receptor的結合，PRMT1的活性會受到調控，而此兩類蛋白質被認為在調控細胞身生長上扮演重要角色，因此我們在RAT1此大鼠纖維母細胞中篩選出過度表現PRMT1的穩定群落，並選取其中一群落65-3，透過計算生長曲線以及BrdU嵌合分析其生長特性，發現其與RAT1細胞在生長上並未有顯著差別。同時，我們針對其細胞粹出物進行活體外甲基化實驗，以測試過度表現PRMT1的酵素活性，結果顯示群落65-3與RAT1細胞中PRMT1的酵素活性並未有明顯差異。

PRMT1 (protein-arginine N-methyltransferase 1) is the predominant arginine methyltransferase in mammalian cells that contributes > 90% of protein-arginine methylated activity. PRMT1 was first cloned because of its ability to interact with mammalian TIS21 immediate-early protein. To understand the enzymatic activity of PRMT1, we used hnRNP A2 as a substrate for GST-PRMT1. We showed that bivalent cations affected the methyltransferase activity of PRMT1. That display distinct methylated degrees in the presences of different cations and concentrations and we found that Ca2+ has conspicuous influence. Relative, univalent cation has slightly effect.
PRMT1 activity appears to be positively regulated through its association with two proteins, the TIS21/BTG1 gene product and the interferon receptor, each of which plays an important role in cell growth regulation. We characterized the growth property of RAT1 cells overexpressing HA-PRMT1 by growth curve counting and BrdU incorporation analysis, and demonstrated that there is no difference between RAT1 and RAT1 overexpressing HA-PRMT1 cells in growth patterns. In addition, in vitro methylation of Trx-hnRNP A2, using cell lysates from RAT1 or RAT1 overexpressing HA-PRMT1, showed that there was no apparent difference in methyltransferases activity.

目 錄 ………………………………………………………………………………Ⅰ
縮 寫 表 ………………………………………………………………………………Ⅱ
圖次目錄 ………………………………………………………………………………Ⅳ
中文摘要 ………………………………………………………………………………1
英文摘要 ………………………………………………………………………………2
緒 論 ………………………………………………………………………………3
研究目標 ………………………………………………………………………………10
實驗材料 ………………………………………………………………………………11
實驗方法 ………………………………………………………………………………19
實驗結果 ………………………………………………………………………………33
討 論 ………………………………………………………………………………43
參考文獻 ………………………………………………………………………………49
圖 ………………………………………………………………………………56
附 圖 ………………………………………………………………………………75

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