質子傳送焦磷酸水解酶是一能維持細胞質內pH恆定的獨特酵素，並藉由水解焦磷酸建立跨生物膜的質子梯度。磷酸化作用隸屬於蛋白質轉譯後修飾的一種，在調控機制中扮演重要的角色。在現階段酵母菌異質體表現綠豆焦磷酸水解酶的研究中，利用Pro-Q 鑽石染漬法發現，焦磷酸水解酶具有磷酸化作用；利用質譜分析法，發現絲胺酸45為磷酸化的位置。除此之外，發現利用酪蛋白激酶2抑制劑，能抑制絲胺酸45的磷酸化作用；第二型蛋白磷酸酶能抑制焦磷酸水解酶的磷酸化作用。綜觀以上結果，提供新的證據顯示酪蛋白激酶2及第二型蛋白磷酸酶，參與調控酵母菌異質表現焦磷酸水解酶磷酸化與去磷酸化作用。

H+-translocating pyrophosphatase (H+-PPases) is a unique enzyme implicated in the cellular pH homeostasis that develops an electrochemical H+ gradient across the membrane through pyrophosphate (PPi) hydrolysis. Phosphorylation is one of the posttranslational modifications (PTMs) of proteins and is considered a regulatory mechanism. In this study, the possible phosphorylation of the Vigna radiata H+-PPases (VrH+-PPases) isolated from the yeast heterologous expression system was revealed for the first time by using the Pro-Q Diamond staining. Furthermore, a phosphorylation site was identified at Ser45 of VrH+-PPases through mass spectroscopy. In addition, the application of casein kinase 2 (CK2) inhibitors resulted in the inhibition of phosphorylation of VrH+-PPases in yeast. Phosphorylation at Ser45 also enhanced the ion effects on VrH+-PPase. On the contrary, the type 2C protein phosphatase (PP2C) was able to dephosphorylate the VrH+-PPase obtained from heterologously expressing yeast. Our study provides new evidence for possible role of CK2 and PP2C in the kinase/phosphatase regulation of VrH+-PPases.

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