TY - JOUR
T1 - An activity stain for dihydroxy-acid dehydratase
AU - Kuo, Che Fu
AU - Mashino, Tadahiko
AU - Fridovich, Irwin
N1 - Funding Information:
’ This work was supported by research grants from the Council for Tobacco Research, U.S.A., Inc.; the United States Army; the National Science Foundation; and the National Institutes of Health.
Copyright:
Copyright 2014 Elsevier B.V., All rights reserved.
PY - 1987/8/1
Y1 - 1987/8/1
N2 - An activity stain has been devised for the dihydroxy-acid dehydratase. When applied to polyacrylamide gel electropherograms of crude soluble extracts of Escherichia coli, it detected a single electromorph. The intensity of staining increased with the amount of extract protein applied to the gel. Activity staining demonstrated that (a) anaerobically grown cells contain more extractable dehydratase activity than do aerobically grown cells; (b) exposure of E. coli to 4.2 atm 02 caused virtually complete loss of activity; (c) exposure of cells to paraquat or plumbagin in the presence of dioxygen, but not in its absence, caused a massive loss of activity. These data illustrate the utility of this activity stain and demonstrate that the dehydratase is inactivated by O2- generated within cells.
AB - An activity stain has been devised for the dihydroxy-acid dehydratase. When applied to polyacrylamide gel electropherograms of crude soluble extracts of Escherichia coli, it detected a single electromorph. The intensity of staining increased with the amount of extract protein applied to the gel. Activity staining demonstrated that (a) anaerobically grown cells contain more extractable dehydratase activity than do aerobically grown cells; (b) exposure of E. coli to 4.2 atm 02 caused virtually complete loss of activity; (c) exposure of cells to paraquat or plumbagin in the presence of dioxygen, but not in its absence, caused a massive loss of activity. These data illustrate the utility of this activity stain and demonstrate that the dehydratase is inactivated by O2- generated within cells.
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U2 - 10.1016/0003-2697(87)90528-8
DO - 10.1016/0003-2697(87)90528-8
M3 - Article
C2 - 2445227
AN - SCOPUS:0023390493
VL - 164
SP - 526
EP - 530
JO - Analytical Biochemistry
JF - Analytical Biochemistry
SN - 0003-2697
IS - 2
ER -