Lichko L P, Kulakovskaia T V, Kulaev I S
Biokhimiia. 1994 Jul;59(7):1088-97.
The effects of some protein kinase effectors on phosphohydrolase and transport activities of yeast vacuoles have been studied. The platelet-activating factor (PAF), a plant vacuolar protein kinase C stimulator, had a protonophoric and membrane-damaging effects on yeast vacuoles and inhibited the ATP-dependent delta mu H+ formation and ATP-dependent secondary transport but stimulated the ATPase and pyrophosphatase hydrolase activities by abrogating proton control. PAF increasing the tonoplast permeability for the corresponding substrates also stimulated pyrophosphatase, polyphosphatase and alkaline phosphatase activities. Lysolipid sphingosine, a plant vacuolar protein kinase C inhibitor, poorly stimulated the ATPase activity and the ATP-dependent formation of Em in isolated yeast vacuoles, while the pyrophosphatase activity increased by 200%. Other hydrolase activities tested were insensitive to the effect of the lysolipid. Sphingosine inhibited the ATP-dependent citrate transport only insignificantly. Heparin, an effective casein kinase inhibitor, suppressed the ATPase and polyphosphatase activities in isolated yeast vacuoles. The polyphosphatase activity was inhibited both in the vacuolar sap and the tonoplast solubilized by a Zwittergent TM-314, in contrast with the ATPase activity which was inhibited by heparin only in isolated vacuoles. Heparin is suggested to inhibit polyphosphatase by directly influencing the enzyme.
已对某些蛋白激酶效应物对酵母液泡磷酸水解酶和转运活性的影响进行了研究。血小板活化因子（PAF），一种植物液泡蛋白激酶C刺激剂，对酵母液泡具有质子载体和膜损伤作用，抑制了ATP依赖的 ΔμH⁺形成和ATP依赖的次级转运，但通过消除质子控制刺激了ATP酶和焦磷酸酶水解酶活性。PAF增加了液泡膜对相应底物的通透性，也刺激了焦磷酸酶、多聚磷酸酶和碱性磷酸酶的活性。溶血脂鞘氨醇，一种植物液泡蛋白激酶C抑制剂，在分离的酵母液泡中对ATP酶活性和Em的ATP依赖形成刺激作用较弱，而焦磷酸酶活性增加了200%。所测试的其他水解酶活性对溶血脂的作用不敏感。鞘氨醇仅轻微抑制了ATP依赖的柠檬酸盐转运。肝素，一种有效的酪蛋白激酶抑制剂，抑制了分离的酵母液泡中的ATP酶和多聚磷酸酶活性。与仅在分离的液泡中被肝素抑制的ATP酶活性相反，多聚磷酸酶活性在液泡液和由两性离子去污剂TM-314增溶的液泡膜中均受到抑制。提示肝素通过直接影响该酶来抑制多聚磷酸酶。
