OppA, the ecto-ATPase of Mycoplasma hominis induces ATP release and cell death in HeLa cells.
OppA, the ecto-ATPase of Mycoplasma hominis induces ATP release and cell death in HeLa cells.
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DOI：
10.1186/1471-2180-8-55
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发表时间：
2008-04-04
期刊：
影响因子：
4.2
通讯作者：
Henrich B
中科院分区：
文献类型：
--
作者：
Hopfe M;Henrich B
在属于无细胞壁柔膜体纲的兼性人类病原体人型支原体中，寡肽通透酶的表面定位底物结合结构域OppA被鉴定为主要的外膜ATP酶。
鉴于细胞外ATP只能由受感染的宿主细胞提供这一观点，我们分析了人型支原体的不同制剂与人宫颈癌细胞（HeLa细胞）孵育后HeLa细胞的ATP释放情况：完整的细菌细胞、含或不含OppA的膜组分、重组OppA以及一种ATP酶缺陷型OppA突变体。在所有缺乏OppA外膜ATP酶活性的样品中，主要测定了ATP释放到HeLa细胞上清液中的情况。在存在ATP酶抑制剂DIDS的情况下，含OppA的样品中ATP的量增加。在用含OppA蛋白的组分孵育后，这种增加达到最大值，这表明OppA参与了ATP释放及随后的水解过程。实时定量PCR分析显示，人型支原体感染后人宫颈癌细胞的增殖减少，流式细胞术实验证实，OppA诱导的人宫颈癌细胞凋亡比坏死更显著，而且OppA外膜ATP酶活性的保留会诱导凋亡。
OppA诱导的人型支原体感染的人宫颈癌细胞的ATP释放和水解所导致的细胞死亡主要是由于凋亡而非坏死。未来的研究将阐明凋亡诱导对于这些非侵入性病原体的生存是否必不可少。
In the facultative human pathogen Mycoplasma hominis, which belongs to the cell wall-less Mollicutes, the surface-localised substrate-binding domain OppA of the oligopeptide permease was characterised as the main ecto-ATPase. With the idea that extra-cellular ATP could only be provided by the infected host cells we analysed the ATP release of HeLa cells after incubation with different preparations of Mycoplasma hominis: intact bacterial cells, the membrane fraction with or without OppA, recombinant OppA as well as an ATPase-deficient OppA mutant. Release of ATP into the supernatant of the HeLa cells was primarily determined in all samples lacking ecto-ATPase activity of OppA. In the presence of the ATPase inhibitor DIDS the amount of ATP in the OppA-containing samples increased. This increase was maximal after incubation with fractions containing OppA protein indicating that OppA is involved in ATP release and subsequent hydrolysis. Real-time PCR analyses revealed that the proliferation of HeLa cells is reduced after infection with M. hominis and flow cytometry experiments established that OppA induces greater apoptosis than necrosis of HeLa cells whereas the preservation of ecto-ATPase activity of OppA induces apoptosis. The OppA induced ATP-release and -hydrolysis induced cell death of M. hominis infected HeLa cells was predominantly due to apoptosis rather than necrosis. Future work will elucidate whether the induction of apoptosis is indispensable for survival of these non-invasive pathogen.
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影响因子：
2.2
作者：
Ferraro, RB;Sousa, JL;Meyer-Fernandes, JR
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DOI：
10.1152/ajpcell.1997.273.3.c973
发表时间：
1997-09-01
影响因子：
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作者：
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