The staphylococcal pep dependent phosphotransferase system: Demonstration of a phosphorylated intermediate of the enzyme 1 component

Stein, Ross L.; Schrecker, Otto; Lauppe, Hans-Frieder; Hengstenberg, H.

doi:10.1016/0014-5793(74)80288-7

Cited by 31 publications

(18 citation statements)

References 9 publications

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“…Evidence for reaction 1, the phosphorylation of Enzyme I by PEP, has recently been presented by Stein et al (12). Other workers have implicated Enzyme I of the PTS sysP tem uniquely in the repression of enzyme synthesis.…”

Section: Discussionmentioning

confidence: 87%

Interaction of enzyme I of the phosphoenolpyruvate:sugar phosphotransferase system with adenylate cyclase of Escherichia coli.

Peterkofsky¹,

Gazdar²

1975

Proc. Natl. Acad. Sci. U.S.A.

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Transient repression by glucose of induced enzyme synthesis involves lowering of intracellular cAMP levels. This glucose effect is partially explained by a glucose inhibition of adenylate cyclase [EC 4.6 (3) supplemented with the designated carbon sources. Strains 1100 (a f-gl+ mutant of Hfr 3000), 1101 (a mutant in the HPr protein of the phosphotransferase system), and 1103 (a leaky mutant in Enzyme I of the PTS system) were originally described by Fox and Wilson (4) and were kindly provided by Dr. Ira Pastan. All assay methods have been previously described. Adenylate cyclase was measured in intact cells by the method of Peterkofsky and Gazdar (5) and in toluene-treated cells by the procedure of Harwood and Peterkofsky (6). Cellular cAMP levels, including a correction for contaminating extracellular cAMP, were determined by the method of Peterkofsky and Gazdar (1). The potassium salt of phosphoenolpyruvate (PEP) was from Calbiochem. RESULTS Abnormal cAMP metabolism in an Enzyme I mutantAn examination was made of the activities of adenylate cyclase in permeabilized cells of various strains of E. coli with lesions in the PTS system. Both strains 1101 (a mutant in the HPr protein) and strain 1103 (a mutant in Enzyme I) were derived from the same parent (strain 1100). The various strains were grown to mid-logarithmic phase on several carbon sources, then tested for adenylate cyclase activity in the presence and absence of glucose. The data of Table 1 show several points of interest. As we previously showed in E. coli B (2), the adenylate cyclase activity can vary depending on the carbon source used for growth. For example, cells of strain 1100 grown on glycerol and glucose-6-phosphate have roughly three times as much activity as cells grown on nutrient broth. Both the wild-type and the HPr mutant show enzyme activities that are inhibited by glucose (1). On the other hand, the Enzyme I mutant shows unusual behavior in two ways. First, irrespective of the carbon source, the activity is substantially lower than that of the other two strains. Second, the activity that is present is not inhibited by glucose.In order to further define the anomalous pattern of adenylate cyclase activity in the Enzyme I mutant, another parameter in cAMP metabolism was measured. Cultures of the three strains were grown in nutrient broth medium to midlogarithmic phase, at which point duplicate samples were taken for determination of intracellular and extracellular cAMP levels. Table 2 shows that the Enzyme I mutant can again be distinguished from the other strains by its low cAMP levels (five to seven times lower intracellular cAMP levels than the other members of the isogenic set).Normal cAMP metabolism of an Enzyme I mutant under carbon-source-starvation conditions A further exploration of the properties of the Enzyme I mutant revealed that the defect in cAMP levels could be re-2920 Abbreviations: PEP, phosphoenolpyruvate; PTS system, phosphoenolpyruvate:sugar phosphotransferase system.

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Section: Discussionmentioning

confidence: 87%

Interaction of enzyme I of the phosphoenolpyruvate:sugar phosphotransferase system with adenylate cyclase of Escherichia coli.

Peterkofsky¹,

Gazdar²

1975

Proc. Natl. Acad. Sci. U.S.A.

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“…Enzyme I: A crude preparation of Enzyme I was used as described previously [3]. The specific activity was about 5 units per mg protein (1 unit = 1 PM orthonitrophenol released in 10 min at 37°C in the extract complementation assay as described earlier) [71.…”

Section: Preparation Of Pi3 Componentsmentioning

confidence: 99%

“…In staphylococcus aureus the PTS has been studied in detail. The following reaction sequence occurs during phosphorylation of galactosides [3]:…”

Section: Introductionmentioning

confidence: 99%

The staphylococcal PEP dependent phosphotransferase system, proton magnetic resonance (PMR) studies on the phosphoryl carrier protein HPr: Evidence for a phosphohistidine residue in the intact phospho‐HPr molecule

Schrecker¹,

Stein²,

Hengstenberg³

et al. 1975

FEBS Letters

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“…The main pleiotropic components are (reaction 3) an Enzyme I that is phosphorylated by PEP at a nitrogen atom of a histidine residue (Stein et al, 1974), and (reaction 4) a small protein, also containing histidine and capable of being phosphorylated at that residue, and designated HPr (histidine-containing protein) for that reason (Kundig et al,I 964).…”

Section: H L Kornbergmentioning

confidence: 99%

Genetics in the Study of Carbohydrate Transport by Bacteria: Sixth Griffith Memorial Lecture

Kornberg

1976

Journal of General Microbiology

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The staphylococcal pep dependent phosphotransferase system: Demonstration of a phosphorylated intermediate of the enzyme 1 component

Cited by 31 publications

References 9 publications

Interaction of enzyme I of the phosphoenolpyruvate:sugar phosphotransferase system with adenylate cyclase of Escherichia coli.

Interaction of enzyme I of the phosphoenolpyruvate:sugar phosphotransferase system with adenylate cyclase of Escherichia coli.

The staphylococcal PEP dependent phosphotransferase system, proton magnetic resonance (PMR) studies on the phosphoryl carrier protein HPr: Evidence for a phosphohistidine residue in the intact phospho‐HPr molecule

Genetics in the Study of Carbohydrate Transport by Bacteria: Sixth Griffith Memorial Lecture

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