Synthese von [<sup>32</sup>P]phosphoenolpyruvat

Lauppe, Hans-Frieder; Rau, Gisela; Hengstenberg, Wolfgang

doi:10.1016/0014-5793(72)80524-6

Cited by 27 publications

(11 citation statements)

References 4 publications

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“…However, this material is not commercially available. Chemical methods such as that described by Lauppe et al (1972) gave variable and low yields in my hands. The enzymic procedure of Kundig & Roseman (1971) is more reproducible, but it requires the use of biological materials, such as coupled mitochondria, which are not always readily available and which cannot be stored without loss of activity.…”

mentioning

confidence: 79%

A simple enzymic method for the synthesis of [32P]phosphoenolpyruvate

Parra

1982

Biochemical Journal

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confidence: 79%

A simple enzymic method for the synthesis of [32P]phosphoenolpyruvate

Parra

1982

Biochemical Journal

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“…Protein phosphorylation. [32P]PEP was synthesized as described previously (7,10). 32P-phosphorylation of enzyme I was performed as follows.…”

Section: Methodsmentioning

confidence: 99%

Evidence for covalently cross-linked dimers and trimers of enzyme I of the Escherichia coli phosphotransferase system

Grenier¹,

Reizer²,

Waygood³

et al. 1985

J Bacteriol

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Enzyme I of the bacterial phosphotransferase system catalyzes transfer of the phosphoryl moiety from phosphoenolpyruvate to both of the heat-stable phosphoryl carrier proteins of the phosphotransferase system, HPr and FPr. Using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and high-pressure liquid chromatography, we demonstrated the existence of covalently cross-linked enzyme I dimers and trimers. Enzyme I exchange assays and phosphorylation experiments with [32PJphosphoenolpyruvate showed that covalent dimers and trimers are catalytically active. Inhibitors of the enzyme I-catalyzed phosphoenolpyruvate-pyruvate exchange block the phosphorylation of enzyme I dimers and trimers. Inhibition of the activity of enzyme I by N-ethylmaleimide, but not that by p-chloromercuriphenylsulfonate, could be overcome by high concentrations of enzyme, suggesting that N-ethylmaleimide modification changes the associative properties of enzyme I. We present evidence for two distinct classes of sulifhydryl groups in enzyme I.The phosphoenolpyruvate (PEP)-dependent sugar phosphotransferase system of bacteria catalyzes the transport and concomitant phosphorylation of a number of sugars (1). One component of the system, enzyme I, catalyzes the two phosphoryl transfer reactions shown below.

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“…3P-labeled phosphoenolpyruvate was synthesized by the method developed by Lauppe et al (13 50 ,uCi), and 10 pl of 500 mM streptozotocin. After 10 min of incubation at room temperature, the cell debris was pelleted by centrifugation.…”

Section: Methodsmentioning

confidence: 99%

“…This in vivo system has the capacity to transfer phosphate residues from the added phosphoenolpyruvate to PTS sugars like amethyl-glucoside (13). We succeeded in phosphorylating streptozotocin with this system.…”

Section: Methodsmentioning

confidence: 99%

Phosphorylation of streptozotocin during uptake via the phosphoenolpyruvate: sugar phosphotransferase system in Escherichia coli

Ammer¹,

Brennenstuhl²,

Schindler³

et al. 1979

Antimicrob Agents Chemother

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Mutants of Escherichia coli K-12, Staphylococcus aureus, and Bacillus subtilis defective in the general components (enzyme I, or HPr, or both) of the phosphoenolpyruvate:sugar phosphotransferase system are shown to be resistant to the antibiotic streptozotocin. It is shown here, employing 32P-labeled phosphoenolpyruvate, that wild-type cells of E. coli phosphorylate streptozotocin, whereas with a phosphotransferase system-defective mutant of E. coli the drug is recovered in an unaltered, free form. The internal accumulation of streptozotocin at the steady-state level was about 70 times that of the concentration in the external medium. The antibacterial action of streptozotocin, as well as the uptake of the drug, was inhibited by N-acetyl-D-glucosamine. The uptake of the antibiotic was extremely sensitive to p-chloromercuribenzoate. It is concluded that streptozotocin is taken up by E. coli via the phosphoenolpyruvate:sugar phosphotransferase system and consequently accumulates in the cell at first as streptozotocin-phosphate.

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Synthese von [³²P]phosphoenolpyruvat

Cited by 27 publications

References 4 publications

A simple enzymic method for the synthesis of [32P]phosphoenolpyruvate

A simple enzymic method for the synthesis of [32P]phosphoenolpyruvate

Evidence for covalently cross-linked dimers and trimers of enzyme I of the Escherichia coli phosphotransferase system

Phosphorylation of streptozotocin during uptake via the phosphoenolpyruvate: sugar phosphotransferase system in Escherichia coli

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Synthese von [32P]phosphoenolpyruvat

Cited by 27 publications

References 4 publications

A simple enzymic method for the synthesis of [32P]phosphoenolpyruvate

A simple enzymic method for the synthesis of [32P]phosphoenolpyruvate

Evidence for covalently cross-linked dimers and trimers of enzyme I of the Escherichia coli phosphotransferase system

Phosphorylation of streptozotocin during uptake via the phosphoenolpyruvate: sugar phosphotransferase system in Escherichia coli

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Synthese von [³²P]phosphoenolpyruvat