title: Production in Escherichia coli of active Sorghum phosphoenolpyruvate carboxylase which can be phosphorylated.
abstract: Phosphoeolpyruvate carboxylase (PEPC)-deficient mutants of Escherichia coli have been complemented with a plasmid bearing a full-length cDNA encoding the C4-type form of Sorghum leaf PEPC. Transformed cells grew on minimal medium. Two clones were selected which produce a functional and full-sized enzyme protein as determined by activity assays, immunochemical behavior and SDS-PAGE. In addition, regulatory phosphorylation of immunopurified recombinant PEPC was observed when the enzyme was incubated with a partially purified plant PEPC kinase. These results establish that E. coli cells produce a genuine, phosphate-free, higher-plant PEPC. Application of immunoadsorbtion chromatography to bacterial extracts makes it possible to prepare highly pure protein available for biochemical studies.
authors.affiliation: Laboratoire de Physiologie Végétale Moléculaire, URA CNRS D 1128, Université de Paris-Sud, Orsay, France.
authors.forename: C
authors.initials: C
authors.lastname: Crétin
authors.forename: N
authors.initials: N
authors.lastname: Bakrim
authors.forename: E
authors.initials: E
authors.lastname: Kéryer
authors.forename: S
authors.initials: S
authors.lastname: Santi
authors.forename: L
authors.initials: L
authors.lastname: Lepiniec
authors.forename: J
authors.initials: J
authors.lastname: Vidal
authors.forename: P
authors.initials: P
authors.lastname: Gadal
mesh_terms: D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D010730:Phosphoenolpyruvate Carboxylase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D010766:Phosphorylation; D010957:Plasmids; D006109:Poaceae / Q000201:enzymology* / Q000235:genetics; D014170:Transformation, Genetic
chemical_list: D010730:Phosphoenolpyruvate Carboxylase