TY - JOUR
T1 - Histidine regulation in Salmonella typhimurium. XVI. A sensitive radiochemical assay for histidinol dehydrogenase
AU - Ciesla, Zygmunt
AU - Salvatore, Francesco
AU - Broach, James R.
AU - Artz, Stanley W.
AU - Ames, Bruce N.
PY - 1975/1
Y1 - 1975/1
N2 - A sensitive radiochemical assay for measurement of histidinol dehydrogenase is presented. The method is based upon separation of the product of the reaction. [14C]histidine, from the substrate, [14C]histidinol, on small Dowex 50 columns. The assay can be performed on cell extracts or on toluenized cells and is approximately 100 times more sensitive than previously reported assays for this enzyme. [14C]histidinol is obtained in high yields through conversion of uniformly labeled 14C-glucose by a strain of Salmonella typhimurium derepressed for the histidine operon and blocked at the histidinol dehydrogenase step. Accumulated [14C]histidinol is purified from the culture supernatant by ion-exchange chromatography. This sensitive assay has facilitated measurement of reduced levels of histidine operon expression in promoter mutants, and has been adapted for study of histidine operon regulation in a cell free protein synthesizing system.
AB - A sensitive radiochemical assay for measurement of histidinol dehydrogenase is presented. The method is based upon separation of the product of the reaction. [14C]histidine, from the substrate, [14C]histidinol, on small Dowex 50 columns. The assay can be performed on cell extracts or on toluenized cells and is approximately 100 times more sensitive than previously reported assays for this enzyme. [14C]histidinol is obtained in high yields through conversion of uniformly labeled 14C-glucose by a strain of Salmonella typhimurium derepressed for the histidine operon and blocked at the histidinol dehydrogenase step. Accumulated [14C]histidinol is purified from the culture supernatant by ion-exchange chromatography. This sensitive assay has facilitated measurement of reduced levels of histidine operon expression in promoter mutants, and has been adapted for study of histidine operon regulation in a cell free protein synthesizing system.
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U2 - 10.1016/0003-2697(75)90187-6
DO - 10.1016/0003-2697(75)90187-6
M3 - Article
C2 - 1089301
AN - SCOPUS:0016427277
SN - 0003-2697
VL - 63
SP - 44
EP - 55
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 1
ER -