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水果果肉中磷酸烯醇式丙酮酸羧激酶(PEPCK)和磷酸烯醇式丙酮酸羧化酶(PEPC)的磷酸化作用

Phosphorylation of phosphoenolpyruvate carboxykinase (PEPCK) and phosphoenolpyruvate carboxylase (PEPC) in the flesh of fruits.

作者信息

Walker Robert P, Paoletti Andrea, Leegood Richard C, Famiani Franco

机构信息

Dipartimento di Scienze Agrarie, Alimentari e Ambientali, Università degli Studi di Perugia, Italy.

Dipartimento di Scienze Agrarie, Alimentari e Ambientali, Università degli Studi di Perugia, Italy.

出版信息

Plant Physiol Biochem. 2016 Nov;108:323-327. doi: 10.1016/j.plaphy.2016.07.021. Epub 2016 Jul 21.

Abstract

This study determined whether phosphoenolpyruvate carboxykinase (PEPCK) and phosphoenolpyruvate carboxylase (PEPC) are phosphorylated in the flesh of a range of fruits. This was done by incubating fruit flesh with P[P] (where P[P] = PO), then PEPCK and PEPC were immunoprecipitated from extracts using specific antisera. The incorporation of P[P] into these enzymes was then determined by autoradiography of SDS-PAGE gels. Both enzymes were subject to phosphorylation in vivo in the flesh of grape, tomato, cherry and plum. PEPCK was also subject to phosphorylation in vivo in developing grape seeds. Proteolytic cleavage of PEPCK showed that it was phosphorylated at a site(s) located on its N-terminal extension. Potentially phosphorylation of these enzymes could contribute to the coordinate regulation of their activities in the flesh of fruits and in developing seeds.

摘要

本研究确定了一系列水果果肉中磷酸烯醇丙酮酸羧激酶(PEPCK)和磷酸烯醇丙酮酸羧化酶(PEPC)是否被磷酸化。通过用P[P](其中P[P]=PO)孵育果肉来完成此操作,然后使用特异性抗血清从提取物中免疫沉淀PEPCK和PEPC。然后通过SDS-PAGE凝胶的放射自显影来确定P[P]掺入这些酶中的情况。在葡萄、番茄、樱桃和李子的果肉中,这两种酶在体内均会发生磷酸化。在发育中的葡萄种子中,PEPCK在体内也会发生磷酸化。PEPCK的蛋白水解切割表明它在其N端延伸部分的一个位点被磷酸化。这些酶的磷酸化可能有助于协调调节它们在水果果肉和发育中的种子中的活性。

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