Respective contribution of mitochondrial superoxide and pH to mitochondria-targeted circularly permuted yellow fluorescent protein (mt-cpYFP) flash activity.
Journal of Biological Chemistry. 2013-04-01; 288(15): 10567-10577
DOI: 10.1074/jbc.m113.455709
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Wei-LaPierre L(1), Gong G, Gerstner BJ, Ducreux S, Yule DI, Pouvreau S, Wang X,
Sheu SS, Cheng H, Dirksen RT, Wang W.
Author information:
(1)Department of Pharmacology and Physiology, University of Rochester Medical
Center, Rochester, New York 14642, USA.
Superoxide flashes are transient bursts of superoxide production within the
mitochondrial matrix that are detected using the superoxide-sensitive biosensor,
mitochondria-targeted circularly permuted YFP (mt-cpYFP). However, due to the pH
sensitivity of mt-cpYFP, flashes were suggested to reflect transient events of
mitochondrial alkalinization. Here, we simultaneously monitored flashes with
mt-cpYFP and mitochondrial pH with carboxy-SNARF-1. In intact cardiac myocytes
and purified skeletal muscle mitochondria, robust mt-cpYFP flashes were
accompanied by only a modest increase in SNARF-1 ratio (corresponding to a pH
increase of