Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/24062
標題: 探討Xanthomonas campestris pv. campestris BCP (bacterioferritin comigratory protein)蛋白質結構及 還原烷基過氧化物的機制
Insights into the alkyl peroxide reduction pathway of Xanthomonas campestris bacterioferritin comigratory protein from the trapped intermediate-ligand complex structures
作者: 廖淑如
Liao, Shu-Ju
關鍵字: peroxide oxidoreduction pathway
過氧化氧化還原酶十字花科黑腐病
BCP
Prxs
atypical 2-Cys
BCP蛋白
非典型2-Cys過氧化氧化還原酶
出版社: 生物化學研究所
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摘要: 需氧生物藉由氧氣以進行體內各項機能的運作,但代謝過程中體內的物質會發生氧化反應而產生過氧化物(peroxides),造成細胞的死亡。因此生物體具有多種清除過氧化物的防禦性酵素,例如:超氧化物歧化酶(superoxide dismutase,簡稱SOD)、過氧化氫酶(Catalase,CAT)、麩胱甘肽過氧化酶(Glutathione peroxidase, GPx)及過氧化氧化還原酶(peroxiredoxins 簡稱 Prxs)。新發現的Prxs氧化還原酶,為硫醇特異性抗氧化蛋白,又稱為thioredoxin peroxidases 及alkyl-hydroperoxide-reductase -C22 proteins。Prxs 依照其所參與氧化還原反應的半胱胺酸(Cysteine)之數目與位置可分為1-Cys Prxs、典型的2-Cys Prxs 及非典型的2-Cys Prxs。Prx 蛋白在不同的生物體中有不同數目的異構型,在哺乳動物細胞中,目前發現有六種(PrxI-VI),而在大腸桿菌中則有三種(AhpC、p20、BCP)。許多典型的2-Cys Prxs 及1-Cys Prxs 的三級結構己經被解析出來,進而瞭解其作用機制。但在非典型的2-Cys Prxs 包含大腸桿菌中之BCP (Bacterioferritin comigratory protein)蛋白的三級結構及作用機制仍不太清楚。在此我們同時解析了XcBCP 氧化態(Intra_SS) ; XcBCP 還原態(Free_SH)及XcBCP 含ligand(Inter-SH)的蛋白晶體結構。分析此三結構的結果指出,其三級結構除了具有高度保留的活化中心結構 ( thioredoxin fold),並進一步發現,其為一新型的非典型2-Cys Prxs 蛋白。並藉由與ligand (DNS)的共結晶,我們可以進一步推測XcBCP蛋白與alkyl peroxide substrates 的作用機制。XcBCP 蛋白藉由N 端的氧化半胱胺酸(peroxidatic cysteine)與C 端的還原半胱胺酸(resolving cysteine),將過氧化物還原。而本身成為具有分子內雙硫鍵的氧化態蛋白,並藉由硫氧化還原蛋(Trx)及其還原酵素(TrxR)將氧化態BCP 蛋白還原成還原態,進而繼續還原下一個過氧化物。
Aerobically growing cells are constantly challenged by reactive oxygen species (ROS), potent oxidants capable of damaging cellular components. To protect against the toxicity of ROS, aerobic organisms are equipped with an array of defensive enzymes, such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and Peroxiredoxin (Prx). Peroxiredoxins (Prxs), also termed the thioredoxin peroxidases and alkyl-hydroperoxide-reductase-C22 protein have received considerable attention in recent years as a new and expanding family of thiol-speific antioxidant proteins. Prxs are classified into three major subfamilies, the 1-Cys, the typical 2-Cys, and the atypical 2-Cys Prx, based on the number and position of Cys residue that participate in catalysis. Many organisms produce more than one isoform of Prx, for examples, mammalian cells express six isoforms of Prx (Prx I to VI) and E coli has three isoforms (AhpC, p20, and BCP). The tertiary structures of several typical 2-Cys and 1-Cys Prx have been determined. However, atypical 2-Cys Prx, including the bacterioferritin comigratory protein (BCP), are less well characterized. Here, we report the crystal structures of XcBCP, a novel member of the atypical 2-Cys Prx from the plant pathogen Xanthomonas campestris, in three different states, the Free_SH state, Intra_SSstate, and Inter _SS state. The oxidoreduction pathway of XcBCP has been obtained from trapped intermediate/ligand complex structures. The XcBCP contains the conserved CP-SH at the N terminus that is oxidized to sulfenic acid (CP-SOH) during the catalytic reaction. The CP-SOH is reduced back to CP-SH by a resolving CR-SH located at the C terminus end of the same subunit to form an intramolecular disulfide bond. The intramolecular disulfide bond of XcBCP can be shuttled back to thiol groups by thioredoxin (XcTrx) to complete the catalytic cycle.
URI: http://hdl.handle.net/11455/24062
其他識別: U0005-2107201016083800
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-2107201016083800
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