Concentration of pyrophosphate in exponential phase on minimal medium

Value 1.3 mM
Organism Bacteria Escherichia coli
Reference Kent RB, Guterman SK. Pyrophosphate inhibition of rho ATPase: a mechanism of coupling to RNA polymerase activity. Proc Natl Acad Sci U S A. 1982 Jul 79(13):3992-6. Table 3PubMed ID6125940
Primary Source Heinonen J. Intracellular concentration of inorganic pyrophosphate in the cells of Escherichia coli: A method for its determination. Anal Biochem. 1974 Jun59(2):366-74PubMed ID4365803
Method (Primary source) First PPi is precipitated as manganous salt. Further purification is achieved in two chromatographic runs performed in the opposite directions on the same sheet of paper. The first solvent system consists of ethyl methyl ketone-methanol-water-36% HCl (20:40:10:1, by vol) and the second of ethyl methyl ketone-methanol-water-trichloroacetic acid-25% NH4-EDTA (40 ml: 10 ml: 15 ml: 5 g: 0.8 ml: 50 mg). Each run takes only about 40 min of time.
Comments Using this procedure the intracellular concentration of PPi in the exponential phase cells of Escherichia coli growing aerobically in minimal medium at 37°C was determined to be about 1.3 mM or 5.5 nmoles/mg of dry weight. See BNID 106780
Entered by Uri M
ID 104792