Abstract
Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and ammonium sulfate fractionation were employed in series to purify and concentrate a 12.5-kDa protein fragment with a periodic (24-min period) proteinase K-resistant and drug-unresponsive NADH oxidase (CNOX) activity from pooled sera from healthy volunteers. The activity was unresponsive to capsaicin to distinguish it from the previously isolated cancer-associated NOX form (tNOX). Polyclonal antisera generated to the CNOX fragment cross-reacted with 20.5- to 24-kDa proteins of human sera, human lymphocytes, and plasma membranes from Escherichia coli with the molecular weight depending on source and conditions of treatment with proteinase K.
MeSH terms
-
Ammonium Sulfate / pharmacology
-
Blotting, Western
-
Capsaicin / pharmacology
-
Cell Fractionation
-
Cell Membrane / chemistry
-
Cell Membrane / metabolism
-
DNA, Complementary / metabolism
-
Disulfides / pharmacology
-
Drug Resistance*
-
Electrophoresis, Polyacrylamide Gel
-
Endopeptidase K / pharmacology
-
Escherichia coli / metabolism
-
Female
-
Humans
-
Male
-
NADH, NADPH Oxidoreductases / chemistry*
-
NADH, NADPH Oxidoreductases / isolation & purification
-
NADH, NADPH Oxidoreductases / metabolism*
-
Pyridines / pharmacology
-
Spectrophotometry
-
Temperature
-
Time Factors
Substances
-
DNA, Complementary
-
Disulfides
-
Pyridines
-
3,3'-dipyridyl disulfide
-
ENOX1 protein, human
-
NADH, NADPH Oxidoreductases
-
Endopeptidase K
-
Capsaicin
-
Ammonium Sulfate