Nicotinamide nucleotide pools 15 min after H2O2 challenge

Range Table - link
Organism Bacteria Escherichia coli
Reference Brumaghim JL, Li Y, Henle E, Linn S. Effects of hydrogen peroxide upon nicotinamide nucleotide metabolism in Escherichia coli: changes in enzyme levels and nicotinamide nucleotide pools and studies of the oxidation of NAD(P)H by Fe(III). J Biol Chem. 2003 Oct 24 278(43):42495-504. p.42498 table IIIPubMed ID12913009
Method "Cells were lysed by NaOH in the presence of cyanide and EDTA. EDTA serves to chelate metal ions that could oxidize NAD(P)H, whereas cyanide forms adducts with NAD(P) to stabilize these species against degradation in basic solution, and these adducts can conveniently be quantitated by absorbance at 327 nm."
Comments "The determination of nicotinamide nucleotide pools is complex because of the sensitivity of NAD(P)H to degradation or oxidation during isolation. Lundquist and Olivera (ref 49) estimated the concentrations of NAD(P) in E. coli but did not include the reduced nicotinamide nucleotides. Bochner and Ames (ref 50) used an acid extraction procedure for purification of the nicotinamide nucleotides which probably resulted in oxidation of NAD(P)H prior to quantitation. Therefore, [investigators] modified the method used for brain cells by Klaidman et al. (ref 37) to measure the effect of oxidative stress upon the free pools in E. coli (Table III)."
Entered by Uri M
ID 108044