Cytochrome P450 enzymes but not NADPH oxidases are the source of the NADPH-dependent lucigenin chemiluminescence in membrane assays

详细信息    查看全文

• 作者：Flá ; via Rezende^a ; Kim-Kristin Prior^a ; Oliver Lö ; we^a ; Ilka Wittig^b ; Valentina Strecker^b ; Franziska Moll^a ; Valeska Helfinger^a ; Frank Schnü ; tgen^c ; Nina Kurrle^c ; Frank Wempe^c ; Maria Walter^a ; Sven Zukunft^d ; Bert Luck^d ; Ingrid Fleming^d ; Norbert Weissmann^e ; Ralf P. Brandes^a ; ^{brandes@vrc.uni-franfurt.de} ; Katrin Schrö ; der^a ; ^{schroeder@vrc.uni-frankfurt.de}
• 关键词：NADPH oxidase ; Nox ; Lucigenin ; Chemiluminescence ; Superoxide ; Reactive oxygen species ; Membrane assays
• 刊名：Free Radical Biology and Medicine
• 出版年：2017
• 出版时间：January 2017
• 年：2017
• 卷：102
• 期：Complete
• 页码：57-66
• 全文大小：874 K
• 卷排序：102

文摘

Nox activity of intact cells could not be recapitulated in membranes treated with NADPH. Proteomics of membranes show P450 reductase as source of NADPH-dependent signal. Microsomes overexpressing Cytochrome P450 system produce a NADPH-dependent signal. Knockout of P450 reductase (CRISPR/Cas9) abolished lucigenin signal in HEK cell membranes. Knockout of POR but not p22phox abolishes the basal and Angiotensin II-stimulated NADPH-dependent signal in SMC membranes.