2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/38036In this project we are going to find out the relationship of protein methylation, mostly protein N-arginine methylation to form symmetric or asymmetric NG, NG –dimethylarginines (sDMA or aDMA), and autoimmune disease. Many methylarginine containing proteins such as fibrillarin, several hnRNP, myelin basic protein and SmD1 and D3 are known to be autoantigens of different autoimmune diseases. Thus in this project we will firstly obtain anti-Sm and anti-RNP autoantibodies from SLE patients and other antisera from the Project Core to search for the methylated autoantigens proteomically. We will perform immunoprecipitation and identify the immunoprecipitated proteins through mass spectrometry and data base searches. Secondly, as apoptosis has been shown to present the proteins inside cells, we will look at the relationship between protein arginine methylation and apoptosis. We will elucidate differences in the protein methylation states upon apoptosis. We will use firstly HeLa cells and then keratinocytes induced by UV-B as well as H2O2 as our apoptosis system. Lastly we will link the relationship between protein methylation modification, apoptosis and autoimmune disease. Thus by the end of the Project, we are expecting to identify the autoantigen patterns of different autoantibodies proteomically and to find out if specifically arginine methylations are present in the proteins and required for the recognition. We will also have elucidated whether apoptosis has any effect on arginine methylation and vice versa. Whether the modification is important to elicit the disease will be answered and whether we can use the knowledge obtained here to help further treatment of the disease will be investigated.在此研究計劃中，我們想要探討蛋白質甲基化和自體免疫疾病的關係，蛋白質甲 基化主要著重於精胺酸的甲基化（分為對稱型或不對稱型? 甲基精胺酸）。許多蛋白質 包含了甲基精胺酸，如：fibrillarin、部分的hnRNP 蛋白、myelin basic protein 以及SmD1、 D3 蛋白，而這些蛋白也已被發現是許多自體免疫疾病的自體免疫抗原。在這個計劃中， 我們首先從紅斑性狼瘡病人中取得anti-Sm和anti-RNP 自體免疫抗體血清及獲取其他的 免疫抗體血清（從Project Core），利用蛋白質體學方法找出有甲基化自體免疫抗原；我 們利用自體免疫抗體血清作免疫沉澱法，免疫沉澱下來的蛋白再作質譜儀分析及資料庫 的? 尋比對。此外，在細胞凋亡的過程中，有許多蛋白會在細胞中呈現；我們想要研究 這些蛋白是否有精胺酸甲基化且甲基化與細胞凋亡之間的關係；也就是探討在細胞凋亡 的過程中，是否蛋白質精胺酸甲基化情況的不同；我們將利用UV-B 或H2O2 誘發HeLa cells 和keratinocytes 來作為細胞凋亡的研究系統。最後，我們將會研究蛋白質甲基化、 細胞凋亡與自體免疫疾病三者之間的關係。利用蛋白質體學方法找出不同自體免疫抗體 所辨識的自體免疫抗原，並看其抗原被辨認的位置是否與精胺酸甲基化有關。以及細胞 凋亡會和精胺酸甲基化之相關性。是否精胺酸甲基化與疾病有直接的關係。期望未來能 將實驗的結果應用在疾病的治療上。Project