TY - JOUR
T1 - A Selective and Cell-Permeable Mitochondrial Calcium Uniporter (MCU) Inhibitor Preserves Mitochondrial Bioenergetics after Hypoxia/Reoxygenation Injury
AU - Woods, Joshua J.
AU - Nemani, Neeharika
AU - Shanmughapriya, Santhanam
AU - Kumar, Akshay
AU - Zhang, Mengqi
AU - Nathan, Sarah R.
AU - Thomas, Manfred
AU - Carvalho, Edmund
AU - Ramachandran, Karthik
AU - Srikantan, Subramanya
AU - Stathopulos, Peter B.
AU - Wilson, Justin J.
AU - Madesh, Muniswamy
N1 - Funding Information:
This research in the laboratory of M. Madesh was funded by the National Institutes of Health (R01GM109882, R01HL086699, R01HL142673, R01HL119306). Research in the laboratory of J. J. Wilson was supported by the National Science Foundation (NSF) (CHE-1750295). S. Shanmugh-apriya is supported by the NIH K99/R00 grant (1 K99 HL138268-01). E. Carvalho and N. Nemani are supported by the AHA fellowships (18POST33990217 and 17PRE33660720). J. J. Woods is supported by the NSF-GRFP (DGE-1650441). This work made use of the Cornell NMR facility, which is funded in part by the NSF (CHE-1531632). We thank Reggie Jacob for helpful comments on the manuscript and Samantha Davalos for assistance in preparing the artwork associated with this manuscript.
Publisher Copyright:
© 2019 American Chemical Society.
PY - 2019/1/23
Y1 - 2019/1/23
N2 - Mitochondrial Ca 2+ ( m Ca 2+ ) uptake mediated by the mitochondrial calcium uniporter (MCU) plays a critical role in signal transduction, bioenergetics, and cell death, and its dysregulation is linked to several human diseases. In this study, we report a new ruthenium complex Ru265 that is cell-permeable, minimally toxic, and highly potent with respect to MCU inhibition. Cells treated with Ru265 show inhibited MCU activity without any effect on cytosolic Ca 2+ dynamics and mitochondrial membrane potential ( m ). Dose-dependent studies reveal that Ru265 is more potent than the currently employed MCU inhibitor Ru360. Site-directed mutagenesis of Cys97 in the N-terminal domain of human MCU ablates the inhibitory activity of Ru265, suggesting that this matrix-residing domain is its target site. Additionally, Ru265 prevented hypoxia/reoxygenation injury and subsequent mitochondrial dysfunction, demonstrating that this new inhibitor is a valuable tool for studying the functional role of the MCU in intact biological models.
AB - Mitochondrial Ca 2+ ( m Ca 2+ ) uptake mediated by the mitochondrial calcium uniporter (MCU) plays a critical role in signal transduction, bioenergetics, and cell death, and its dysregulation is linked to several human diseases. In this study, we report a new ruthenium complex Ru265 that is cell-permeable, minimally toxic, and highly potent with respect to MCU inhibition. Cells treated with Ru265 show inhibited MCU activity without any effect on cytosolic Ca 2+ dynamics and mitochondrial membrane potential ( m ). Dose-dependent studies reveal that Ru265 is more potent than the currently employed MCU inhibitor Ru360. Site-directed mutagenesis of Cys97 in the N-terminal domain of human MCU ablates the inhibitory activity of Ru265, suggesting that this matrix-residing domain is its target site. Additionally, Ru265 prevented hypoxia/reoxygenation injury and subsequent mitochondrial dysfunction, demonstrating that this new inhibitor is a valuable tool for studying the functional role of the MCU in intact biological models.
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U2 - 10.1021/acscentsci.8b00773
DO - 10.1021/acscentsci.8b00773
M3 - Article
AN - SCOPUS:85059762454
SN - 2374-7943
VL - 5
SP - 153
EP - 166
JO - ACS Central Science
JF - ACS Central Science
IS - 1
ER -