Decreased protein nitration in macrophages that overexpress indoleamine 2, 3-dioxygenase.
Decreased protein nitration in macrophages that overexpress indoleamine 2, 3-dioxygenase.
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DOI：
10.2478/s11658-006-0048-9
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发表时间：
2007
影响因子：
8.3
通讯作者：
Gearhart DA
中科院分区：
文献类型：
--
作者：
Keskin DB;Marshall B;Munn D;Mellor AL;Gearhart DA
吲哚胺2，3-双加氧酶（IDO; E.C. 1.13.11.42）催化色氨酸的氧化裂解形成犬尿氨酸。IDO活性消耗超氧阴离子;因此，我们推测IDO的过表达可能减轻体外细胞蛋白质的超氧阴离子依赖性氧化修饰。我们制备并表征了用IDO表达载体或对照（空）载体稳定转染的RAW 264.7巨噬细胞。我们在IDO转染的巨噬细胞中检测到IDO mRNA、蛋白和酶活性，但在用空载体转染的巨噬细胞中未检测到。为了原位产生超氧阴离子，我们用黄嘌呤或次黄嘌呤处理IDO和对照转染培养物，然后使用ELISA方法定量总细胞裂解物中氧化修饰蛋白的相对水平。蛋白质羰基的水平在IDO转染和载体转染的巨噬细胞中相似;然而，与对照转染子相比，IDO转染细胞中的蛋白质硝化显著减少。此外，超氧阴离子的稳态水平显着低于IDO转染的文化与对照转染。我们的研究结果是一致的概念，除了降解色氨酸，IDO活性可以保护细胞免受氧化损伤。
The activity of indoleamine 2, 3-dioxygenase (IDO; E.C. 1.13.11.42) catalyzes the oxidative cleavage of tryptophan to form kynurenine. IDO activity consumes superoxide anions; therefore, we postulated that over-expression of IDO might mitigate superoxide-anion dependent, oxidative modification of cellular proteins in vitro. We prepared and characterized RAW 264.7 macrophages that were stably transfected with either an IDO expression vector or the control (empty) vector. We detected IDO mRNA, protein, and enzyme activity in the IDO-transfected macrophages, but not in the macrophages transfected with the empty vector. To generate superoxide anions in situ, we treated the IDO-and control-transfected cultures with xanthine or hypoxanthine, and then used ELISA methods to quantitate the relative levels of oxidatively modified proteins in total cell lysates. The levels of protein carbonyls were similar in IDO-transfected and vector-transfected macrophages; however, protein nitration was significantly less in IDO-transfected cells compared to control transfectants. In addition, steady-state levels of superoxide anions were significantly lower in the IDO-transfected cultures compared with control transfectants. Our results are consistent with the concept that, besides degrading tryptophan, IDO activity may protect cells from oxidative damage.
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影响因子：
3.1
作者：
GUPTA, SL;CARLIN, JM;MURPHY, MJ
通讯作者：
MURPHY, MJ
影响因子：
3.4
作者：
Contestabile, Antonio
通讯作者：
Contestabile, Antonio
影响因子：
9.8
作者：
Betteridge, DJ
通讯作者：
Betteridge, DJ
影响因子：
3.9
作者：
Grant, RS;Passey, R;Kapoor, V
通讯作者：
Kapoor, V
影响因子：
7.4
作者：
Buss, H;Chan, TP;Winterbourn, CC
通讯作者：
Winterbourn, CC