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比较HeLa(癌细胞)和CHO(非癌细胞)细胞的耗氧量的周期性波动以及对外部NAD(P)+/NAD(P)H的反应。

Periodic fluctuations in oxygen consumption comparing HeLa (cancer) and CHO (non-cancer) cells and response to external NAD(P)+/NAD(P)H.

作者信息

Orczyk John, Morré Dorothy M, Morré D James

机构信息

Department of Medicinal Chemistry, Purdue University, West Lafayette, Indiana 47907-2064, USA.

出版信息

Mol Cell Biochem. 2005 May;273(1-2):161-7. doi: 10.1007/s11010-005-0326-2.

Abstract

Oxygen consumption in the presence of cyanide was utilized as a measure of plasma membrane electron transport in Chinese hamster ovary (CHO) and human cervical carcinoma (HeLa) cell lines. Both intact cells and isolated plasma membranes carry cyanide-insensitive NADH(P)H oxidases at their external membrane surfaces (designated ECTO-NOX proteins). Regular oscillatory patterns of oxygen consumption with period lengths characteristic of those observed for rates of NADH oxidation by ECTO-NOX proteins were observed to provide evidence for transfer of protons and electrons to reduce oxygen to water. The oscillations plus the resistance to inhibition by cyanide identify the bulk of the oxygen consumption as due to ECTO-NOX proteins. With intact CHO cells, oxygen consumption was enhanced by but not dependent upon external NAD(P)H addition. With intact HeLa cells, oxygen consumption was inhibited by both NADH and NAD+ as was growth. The results suggest that plasma membrane electron transport from internal donors to oxygen as an external acceptor is mediated through ECTO-NOX proteins and that electron transport to molecular oxygen may be differentially affected by external pyridine nucleotides depending on cell type.

摘要

在存在氰化物的情况下,利用氧消耗来衡量中国仓鼠卵巢(CHO)细胞系和人宫颈癌(HeLa)细胞系中的质膜电子传递。完整细胞和分离的质膜在其外膜表面均携带对氰化物不敏感的NADH(P)H氧化酶(称为ECTO-NOX蛋白)。观察到氧消耗的规则振荡模式,其周期长度与ECTO-NOX蛋白氧化NADH的速率所观察到的特征一致,这为质子和电子转移以将氧还原为水提供了证据。这些振荡加上对氰化物抑制的抗性表明,大部分氧消耗是由ECTO-NOX蛋白引起的。对于完整的CHO细胞,外部添加NAD(P)H可增强氧消耗,但并非依赖于此。对于完整的HeLa细胞,NADH和NAD + 均抑制氧消耗以及细胞生长。结果表明,从内部供体到作为外部受体的氧的质膜电子传递是通过ECTO-NOX蛋白介导的,并且根据细胞类型,向分子氧的电子传递可能受到外部吡啶核苷酸的不同影响。

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