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題 名 | An Essential Arginine Residue in Vacuolar H狇-ATPase Purified from Etiolated Mung Bean Seedlings=白化綠豆幼苗液泡質子傳送核苷三磷酸水解酶中之基要精氨酸 |
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作 者 | 郭順宇; 曾驥孟; 林文傑; 江士昇; 洪淑嫻; 潘榮隆; | 書刊名 | Botanical Bulletin of Academia Sinica |
卷 期 | 40:2 1999.04[民88.04] |
頁 次 | 頁127-134 |
分類號 | 341.94 |
關鍵詞 | H狇-ATPase; 液泡; 質子傳送; 核苷三磷酸水解酶; 化學修飾; 綠豆; Chemical modification; Mung bean; Proton translocation; Tonoplast; |
語 文 | 英文(English) |
中文摘要 | 由白化綠豆幼苗抽出之液泡質子核甘三磷酸水解酯 , 經苯基乙二醛 (Phenylglyoxa1) 或2,3-丁烯雙酮 (2,3-Butanedione) 之處理 , 會造成核甘三磷酸水解作用與質子傳送作用的抑制現象。動力學分析得知 , 苯基乙二醛對液泡質子核甘三磷酸水解抑制之一級反應常數 (k2) 與穩態解離常數 (K1) 分別為0.416s[] 與193mM; 而2,3-丁烯雙酮對液泡質子核甘三磷酸水解抑制之一級反應常數 (k[]) 與穩態解離常數 (K[]) 則分別為0.227s[] 與24.2mM 。同時 ,苯基乙二醛與丁烯雙酮對液泡質子核甘三磷酸水解抑制之級數為0.94與0.89, 指證出液泡質子核甘三磷酸水解[]中至少有一精氯酸被此兩化學物所修飾。李-博氏作圖 (Lineweaver-Burkplot) 顯示此二修飾物之抑制作用為競爭性反應。液泡質子核甘三磷酸水解?的基質鑽入 核甘三磷酸可以有效的保護苯基乙二醛與2,3-.T丁烯雙酮對液泡質子核甘三磷酸水解?之抑制作用。因此, 我們推論植物液泡質子核甘三磷酸水解[]反應中心具有一基要精氯酸 , 此一精氯酸參與基質之結合作用 , 並且促使苯基乙二醛與2,3-丁烯雙酮對液泡質子核甘三磷酸水解[]之抑制作用。 |
英文摘要 | Treatments of the tonoplast ATPase purified from mung bean seedlings (Vigna radiate L.) with guanidino modifiers, phenylglyoxal and 2,3-butanedione, caused a marked loss of the ATP hydrolysis activity and proton translocation in a concentration-dependent manner. Kinetic analysis yielded first order rate constants, k,, of 0.416 and 0.227 s-' and steady-state dissociation constants, K., of 19.3 and 24.2 mM for phenylglyoxal- and butanedione-inhibition of vacuolar H`-ATPase, respectively. The reaction order of phenylglyoxal- and butanedione-inhibition was calculated to be 0.94 and 0.89, respectively, suggesting that at least one arginine residue of vacuolar H'-ATPase was modified by both reagents. Lineweaver-Burk plots showed that the mode of inhibition of vacuolar H ATPase by both modifiers is competitive. Mg-ATP, the physiological substrate of vacuolar H`-ATPase, but not its analogs, exerted preferentially partial protection against phenylglyoxal and butanedione, indicating that the arginine residue involved in the inhibition of enzymatic activity may be located at or near the active site and directly participate in the binding of the substrate. |
本系統中英文摘要資訊取自各篇刊載內容。