Pacquit V, Santi S, Cretin C, Bui V L, Vidal J, Gadal P
Laboratoire de Physiologie Végétale Moléculaire, Unité de Recherche Associée CNRS D-1128, Université de Paris-Sud, Orsay, France.
Biochem Biophys Res Commun. 1993 Dec 30;197(3):1415-23. doi: 10.1006/bbrc.1993.2635.
In this work, the C3-type form of Sorghum phosphoenolpyruvate carboxylase (PyrPC) was produced in PyrPC-deficient strains of Escherichia coli transformed by a plasmid bearing the corresponding full-length cDNA (CPR1). The full-sized protein was purified to homogeneity by immunoaffinity chromatography. Some functional and regulatory properties were described; notably, the immunopurified PyrPC could be phosphorylated in reconstituted assay by 1) both a mammalian PKA and the PyrPC protein serine kinase purified from Sorghum leaves and 2) a novel protein kinase affinity-purified from Sorghum roots. In all cases phosphorylation was accompanied by a marked reduction in its malate sensitivity.
在本研究中，通过携带相应全长cDNA（CPR1）的质粒转化的磷酸烯醇式丙酮酸羧化酶（PyrPC）基因缺失的大肠杆菌菌株中产生了高粱磷酸烯醇式丙酮酸羧化酶（PyrPC）的C3型形式。通过免疫亲和层析将全尺寸蛋白质纯化至同质。描述了一些功能和调节特性；值得注意的是，免疫纯化的PyrPC在重构测定中可被以下物质磷酸化：1）哺乳动物蛋白激酶A和从高粱叶片中纯化的PyrPC蛋白丝氨酸激酶；2）从高粱根中亲和纯化的一种新型蛋白激酶。在所有情况下，磷酸化都伴随着其对苹果酸敏感性的显著降低。
