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首页> 外文期刊>American Journal of Physiology >Anion channels transport ATP into the Golgi lumen.
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Anion channels transport ATP into the Golgi lumen.

机译：阴离子通道将ATP转运到高尔基体腔中。

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Anion channels provide a pathway for Cl(-) influx into the lumen of the Golgi cisternae. This influx permits luminal acidification by the organelle's H(+)-ATPase. Three different experimental approaches, electrophysiological, biochemical, and proteomic, demonstrated that two Golgi anion channels, GOLAC-1 and GOLAC-2, also mediate ATP anion transport into the Golgi lumen. First, GOLAC-1 and -2 were incorporated into planar lipid bilayers, and single-channel recordings were obtained. Low ionic activities of K(2)ATP added to the cis-chamber directly inhibited the Cl(-) subconductance levels of both channels, with K(m) values ranging from 16 to 115 microM. Substitution of either K(2)ATP or MgATP for Cl(-) on the cis, trans, or both sides indicated that ATP is conducted by the channels with a relative permeability sequence of Cl(-) > ATP(4-) > MgATP(2-). Single-channel currents were observed at physiological concentrations of Cl(-) and ATP, providing evidence for their importance in vivo. Second, transport of [alpha-(32)P]ATP into sealed Golgi vesicles that maintain in situ orientation was consistent with movement through the GOLACs because it exhibited little temperature dependence and was saturated with an apparent K(m) = 25 microM. Finally, after transport of [gamma-(32)P]ATP, a protease-protection assay demonstrated that proteins are phosphorylated within the Golgi lumen, and after SDS-PAGE, the proteins in the phosphorylated bands were identified by mass spectrometry. GOLAC conductances, [alpha-(32)P]ATP transport, and protein phosphorylation have identical pharmacological profiles. We conclude that the GOLACs play dual roles in the Golgi complex, providing pathways for Cl(-) and ATP influx into the Golgi lumen.
机译：阴离子通道为Cl（-）流入高尔基池的内腔提供了途径。这种涌入允许通过细胞器的H（+）-ATPase进行腔内酸化。三种不同的实验方法（电生理，生化和蛋白质组学）表明，两个高尔基阴离子通道GOLAC-1和GOLAC-2也介导ATP阴离子转运到高尔基体腔中。首先，将GOLAC-1和-2掺入平面脂质双层中，并获得单通道记录。 K（2）ATP添加到顺式腔室的低离子活性直接抑制了两个通道的Cl（-）亚导水平，K（m）值范围从16到115 microM。用K（2）ATP或MgATP替代顺式，反式或两侧的Cl（-）表示ATP由相对渗透性序列为Cl（-）> ATP（4-）> MgATP的通道进行（2-）。在Cl（-）和ATP的生理浓度下观察到单通道电流，为其在体内的重要性提供了证据。其次，将[α-（32）P] ATP转运到保持原位定向的密封高尔基囊泡中与通过GOLAC的运动一致，因为它对温度的依赖性很小，并且被表观K（m）= 25 microM饱和。最后，在运输了[γ-（32）P] ATP之后，蛋白酶保护实验表明蛋白质在高尔基体腔内被磷酸化，在SDS-PAGE后，通过质谱鉴定了磷酸化条带中的蛋白质。 GOLAC电导，[α-（32）P] ATP转运和蛋白质磷酸化具有相同的药理学特征。我们得出的结论是，GOLAC在高尔基体中扮演双重角色，为Cl（-）和ATP流入高尔基体腔提供了途径。

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