TY - JOUR
T1 - Monitoring methionine sulfoxide with stereospecific mechanism-based fluorescent sensors
AU - Tarrago, Lionel
AU - Péterfi, Zalán
AU - Lee, Byung Cheon
AU - Michel, Thomas
AU - Gladyshev, Vadim N.
N1 - Funding Information:
We thank P. Rey (Laboratoire d’Ecophysiologie Moléculaire des Plantes, UMR7265 CEA-CNRS-Aix-Marseille Université) for the kind gift of dabsyl-MetO and V. Belousov and V. Verkhusha for discussion. This study was supported by US National Institutes of Health grants AG021518 and GM065204 to V.N.G. and HL48743 to T.M.
Publisher Copyright:
© 2015 Nature America, Inc. All rights reserved.
PY - 2015/5/1
Y1 - 2015/5/1
N2 - Methionine can be reversibly oxidized to methionine sulfoxide (MetO) under physiological and pathophysiological conditions, but its use as a redox marker suffers from the lack of tools to detect and quantify MetO within cells. In this work, we created a pair of complementary stereospecific genetically encoded mechanism-based ratiometric fluorescent sensors of MetO by inserting a circularly permuted yellow fluorescent protein between yeast methionine sulfoxide reductases and thioredoxins. The two sensors, respectively named MetSOx and MetROx for their ability to detect S and R forms of MetO, were used for targeted analysis of protein oxidation, regulation and repair as well as for monitoring MetO in bacterial and mammalian cells, analyzing compartment-specific changes in MetO and examining responses to physiological stimuli.
AB - Methionine can be reversibly oxidized to methionine sulfoxide (MetO) under physiological and pathophysiological conditions, but its use as a redox marker suffers from the lack of tools to detect and quantify MetO within cells. In this work, we created a pair of complementary stereospecific genetically encoded mechanism-based ratiometric fluorescent sensors of MetO by inserting a circularly permuted yellow fluorescent protein between yeast methionine sulfoxide reductases and thioredoxins. The two sensors, respectively named MetSOx and MetROx for their ability to detect S and R forms of MetO, were used for targeted analysis of protein oxidation, regulation and repair as well as for monitoring MetO in bacterial and mammalian cells, analyzing compartment-specific changes in MetO and examining responses to physiological stimuli.
UR - http://www.scopus.com/inward/record.url?scp=84940005486&partnerID=8YFLogxK
U2 - 10.1038/nchembio.1787
DO - 10.1038/nchembio.1787
M3 - Article
C2 - 25799144
AN - SCOPUS:84940005486
VL - 11
SP - 332
EP - 338
JO - Nature Chemical Biology
JF - Nature Chemical Biology
SN - 1552-4450
IS - 5
ER -