粒線體內肝醣合成酶激酶 3beta 蛋白質複合 體之蛋白質體研究

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粒線體內肝醣合成酶激酶 3beta 蛋白質複合 體之蛋白質體研究

肝醣合成酶激酶 3beta ,被認為是調控細胞內糖類代謝的關鍵性激酶。近來在

研究上發現到肝醣合成酶激酶 3beta 在細胞受到刺激時會進入粒線體中,而粒 線體在細胞的能量代謝及生死調控扮演了無比重要的角色,但肝醣合成酶激 酶 3beta 於粒線體中執行何種功能並未被研究闡明。由於粒線體中的蛋白質都 以蛋白質複合體的形式存在,所以本研究假設肝醣合成酶激酶 3beta 於粒線體 內亦是以蛋白質複合體的形式存在。所以實驗設計利用免疫沉澱法結合二維 電泳分離粒線體中與肝醣合成酶激酶 3beta 交互作用的蛋白質,在二維電泳膠 片上發現到 28 個蛋白質點;而後利用基質輔助雷射吸附飛行時間質譜儀分析 鑑別這些蛋白質,經資料庫的分析比對後整理出 12 個蛋白質分子。 4 個蛋白 質是參與於代謝路徑及克氏循環中的酵素為 glyceraldehydes-3-phosphate dehyd rogenase, isocitrate dehydrogenase, glutamine synthetase, 和 dihydrolipoyl dehydr ogenase ( pyruvate dehydrogenase subunit ) 4 個蛋白質點是電子傳遞鏈氧化 磷酸化複合體 NADH-ubiquinone oxidoreductase MLRQ subunit (complex I) , ATP synthase-alpha, beta and O subunit (complex V) 。而最後 4 個蛋白質為被報 導過參與細胞生死調控與粒線體增生的蛋白 voltage-dependent anion-selective c hannel 1 (VDAC-1) , mitochondrial elongation factor Tu , cofilin 和 prohibitin

;綜合以上的發現可以為繼續去探索肝醣合成酶激酶 3beta 調控細胞能量代謝 與細胞凋亡的機制提供更多線索。

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Proteomic analysis of mitochondrial proteins form complex with glycogen synthase kinase

3beta

Glycogen synthanse kinase 3beta(GSK3-beta), a kinase considered to mediate gluc ose metabolism in cytosol of the cell, was found can be stimulated to translocate to mitochondria, but how GSK3-beta exerts its effects in mitochondria remain unclear . Mitochondria play a key role in regulating cell life. For exploring mitochondrial p roteins which complex with GSK3-beta, we used immunoprecipitation, two-dimens ional electrophoresis and matrix-assisted laser desorption/ionization mass spectrom etry (MALDI-TOF MS) to identify proteins spots of the 2D map. More than 31 prot ein spots were found co-precipitated with anti-GSK3-betaantibodies, 12 of which w ere identified by MALDI-TOF MS or MALDI-Q-TOF MS/MS. Among the 12 prot eins, four proteins involved in metabolism and the Krebs cycle in mitochondria. Th ey were identified as glyceraldehydes-3-phosphate dehydrogenase, isocitrate dehyd rogenase 3 (NAD+)-3 , glutamine synthetase, and dihydrolipoyl dehydrogenase. T  he other four proteins are involved in electron transfer complex I and V. They are NADH-ubiquinone oxidoreductase MLRQ subunit (complex I), ATP synthase-alph a, beta and O subunit (complex V). The last four proteins, voltage-dependent anion- selective channel 1 (VDAC-1), mitochondrial elongation factor Tu, cofilin, and pro hibitin. These findings might provide clues for understanding the mechanism of ho w GSK3-beta to regulate the metabolism and apoptosis of the cell.

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