Journal
BIOCHEMICAL JOURNAL
Volume 480, Issue 5, Pages 363-384Publisher
PORTLAND PRESS LTD
DOI: 10.1042/BCJ20220611
Keywords
-
Categories
Ask authors/readers for more resources
Superoxide/hydrogen peroxide production by site IQ in complex I occurs during both reverse and forward electron transport, and is equally sensitive to S1QELs, rotenone, and piericidin A. A new assay is introduced to determine the direction of electron flow through complex I. The results show that superoxide/hydrogen peroxide production by site IQ in cells is S1QEL-sensitive and occurs during forward electron transport.
Superoxide/hydrogen peroxide production by site IQ in complex I of the electron transport chain is conventionally assayed during reverse electron transport (RET) from ubiquinol to NAD. However, S1QELs (specific suppressors of superoxide/hydrogen peroxide produc-tion by site IQ) have potent effects in cells and in vivo during presumed forward electron transport (FET). Therefore, we tested whether site IQ generates S1QEL-sensitive super-oxide/hydrogen peroxide during FET (site IQf), or alternatively, whether RET and asso-ciated S1QEL-sensitive superoxide/hydrogen peroxide production (site IQr) occurs in cells under normal conditions. We introduce an assay to determine if electron flow through complex I is thermodynamically forward or reverse: on blocking electron flow through complex I, the endogenous matrix NAD pool will become more reduced if flow before the challenge was forward, but more oxidised if flow was reverse. Using this assay we show in the model system of isolated rat skeletal muscle mitochondria that superoxide/hydro-gen peroxide production by site IQ can be equally great whether RET or FET is running. We show that sites IQr and IQf are equally sensitive to S1QELs, and to rotenone and pieri-cidin A, inhibitors that block the Q-site of complex I. We exclude the possibility that some sub-fraction of the mitochondrial population running site IQr during FET is responsible for S1QEL-sensitive superoxide/hydrogen peroxide production by site IQ. Finally, we show that superoxide/hydrogen peroxide production by site IQ in cells occurs during FET, and is S1QEL-sensitive.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available