title: Phosphorylation of Escherichia coli enolase.
abstract: In vivo labeling of Escherichia coli JA200 pLC 11-8 resulted in 32P incorporation into enolase as demonstrated by immunoaffinity chromatography and electrophoresis followed by autoradiography. Complete acid hydrolysis, followed by thin layer chromatography was employed for determination of the phosphoamino acid residue. Comparison with phosphoamino acid standards resulted in the identification of a labeled residue corresponding to phosphoserine. In vitro labeling of cell extracts from glucose and acetate grown cells resulted in differential labeling of enolase. When specific radioactivities of in vivo labeled enolase were compared, 7 times more label was incorporated at late log phase in glucose grown cells than in late log acetate grown cells. At stationary phase, only 2.5 times more label was incorporated into glucose compared to acetate. When 32P-labeled enolase from glucose grown cells was subjected to treatment with potato acid phosphatase, dephosphorylation of the enzyme could be observed. Monitoring enzyme activity during the acid phosphatase treatment revealed a 70% decrease for the forward enzyme reaction, and a 3-fold increase, followed by a gradual decrease to almost zero, for the reverse enzyme reaction. Complete reversal of the changes in activity was possible by adding an aliquot of partially purified enolase kinase plus ATP.
authors.affiliation: Department of Microbiology, Arizona State University, Tempe 85287.
authors.forename: H K
authors.initials: HK
authors.lastname: Dannelly
authors.forename: B
authors.initials: B
authors.lastname: Duclos
authors.forename: A J
authors.initials: AJ
authors.lastname: Cozzone
authors.forename: H C
authors.initials: HC
authors.lastname: Reeves
mesh_terms: D000135:Acid Phosphatase / Q000493:pharmacokinetics / Q000494:pharmacology; D001345:Autoradiography; D015153:Blotting, Western; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000201:enzymology*; D010759:Phosphorus Isotopes; D010766:Phosphorylation
chemical_list: D010759:Phosphorus Isotopes; D000135:Acid Phosphatase