A drug-unresponsive and protease-resistant CNOX protein from human sera.
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.
Sedlak
D
D
Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University, West Lafayette, Indiana 47907-1333, USA.
Mooré
D M
DM
Mooré
D J
DJ
D000645:Ammonium Sulfate / Q000494:pharmacology; D015153:Blotting, Western; D002211:Capsaicin / Q000494:pharmacology; D002458:Cell Fractionation; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D004220:Disulfides / Q000494:pharmacology; D004351:Drug Resistance; D004591:Electrophoresis, Polyacrylamide Gel; D019286:Endopeptidase K / Q000494:pharmacology; D004926:Escherichia coli / Q000378:metabolism; D005260:Female; D006801:Humans; D008297:Male; D009247:NADH, NADPH Oxidoreductases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D011725:Pyridines / Q000494:pharmacology; D013053:Spectrophotometry; D013696:Temperature; D013997:Time Factors
D018076:DNA, Complementary; D004220:Disulfides; D011725:Pyridines; C004847:3,3'-dipyridyl disulfide; C427863:ENOX1 protein, human; D009247:NADH, NADPH Oxidoreductases; D019286:Endopeptidase K; D002211:Capsaicin; D000645:Ammonium Sulfate