Novel Inhibitors of Mitochondrial Sn-glycerol 3-phosphate Dehydrogenase

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2014 Mar 4

PMID 24587137

Citations 30

Authors

Adam L Orr

Deepthi Ashok

Melissa R Sarantos

Ryan Ng

Tong Shi

Akos A Gerencser

Robert E Hughes

Martin D Brand

Affiliations

Soon will be listed here.

Abstract

Mitochondrial sn-glycerol 3-phosphate dehydrogenase (mGPDH) is a ubiquinone-linked enzyme in the mitochondrial inner membrane best characterized as part of the glycerol phosphate shuttle that transfers reducing equivalents from cytosolic NADH into the mitochondrial electron transport chain. Despite the widespread expression of mGPDH and the availability of mGPDH-null mice, the physiological role of this enzyme remains poorly defined in many tissues, likely because of compensatory pathways for cytosolic regeneration of NAD⁺ and mechanisms for glycerol phosphate metabolism. Here we describe a novel class of cell-permeant small-molecule inhibitors of mGPDH (iGP) discovered through small-molecule screening. Structure-activity analysis identified a core benzimidazole-phenyl-succinamide structure as being essential to inhibition of mGPDH while modifications to the benzimidazole ring system modulated both potency and off-target effects. Live-cell imaging provided evidence that iGPs penetrate cellular membranes. Two compounds (iGP-1 and iGP-5) were characterized further to determine potency and selectivity and found to be mixed inhibitors with IC₅₀ and K(i) values between ∼1-15 µM. These novel mGPDH inhibitors are unique tools to investigate the role of glycerol 3-phosphate metabolism in both isolated and intact systems.

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References

Grimmsmann T, Rustenbeck I . Direct effects of diazoxide on mitochondria in pancreatic B-cells and on isolated liver mitochondria. Br J Pharmacol. 1998; 123(5):781-8. PMC: 1565223. DOI: 10.1038/sj.bjp.0701663. View

Weitzel J, Kutz S, Radtke C, Grott S, Seitz H . Hormonal regulation of multiple promoters of the rat mitochondrial glycerol-3-phosphate dehydrogenase gene: identification of a complex hormone-response element in the ubiquitous promoter B. Eur J Biochem. 2001; 268(14):4095-103. DOI: 10.1046/j.1432-1327.2001.02332.x. View

Choi S, Gerencser A, Nicholls D . Bioenergetic analysis of isolated cerebrocortical nerve terminals on a microgram scale: spare respiratory capacity and stochastic mitochondrial failure. J Neurochem. 2009; 109(4):1179-91. PMC: 2696043. DOI: 10.1111/j.1471-4159.2009.06055.x. View

Cooney G, Dawson A . Effects of indomethacin on respiration and the alpha-glycerolphosphate shuttle in rat kidney mitochondria. Biochem Pharmacol. 1979; 28(7):1067-70. DOI: 10.1016/0006-2952(79)90305-8. View

Koza R, Kozak U, Brown L, Leiter E, MacDonald M, Kozak L . Sequence and tissue-dependent RNA expression of mouse FAD-linked glycerol-3-phosphate dehydrogenase. Arch Biochem Biophys. 1996; 336(1):97-104. DOI: 10.1006/abbi.1996.0536. View

Schryvers A, Lohmeier E, Weiner J . Chemical and functional properties of the native and reconstituted forms of the membrane-bound, aerobic glycerol-3-phosphate dehydrogenase of Escherichia coli. J Biol Chem. 1978; 253(3):783-8. View

Klingenberg M . Localization of the glycerol-phosphate dehydrogenase in the outer phase of the mitochondrial inner membrane. Eur J Biochem. 1970; 13(2):247-52. DOI: 10.1111/j.1432-1033.1970.tb00924.x. View

Brideau C, Gunter B, Pikounis B, Liaw A . Improved statistical methods for hit selection in high-throughput screening. J Biomol Screen. 2004; 8(6):634-47. DOI: 10.1177/1087057103258285. View

Garrib A, McMurray W . Purification and characterization of glycerol-3-phosphate dehydrogenase (flavin-linked) from rat liver mitochondria. J Biol Chem. 1986; 261(17):8042-8. View

10.

Affourtit C, Quinlan C, Brand M . Measurement of proton leak and electron leak in isolated mitochondria. Methods Mol Biol. 2011; 810:165-82. DOI: 10.1007/978-1-61779-382-0_11. View

11.

Quinlan C, Gerencser A, Treberg J, Brand M . The mechanism of superoxide production by the antimycin-inhibited mitochondrial Q-cycle. J Biol Chem. 2011; 286(36):31361-72. PMC: 3173136. DOI: 10.1074/jbc.M111.267898. View

12.

Rauchova H, Drahota Z, Rauch P, Fato R, Lenaz G . Coenzyme Q releases the inhibitory effect of free fatty acids on mitochondrial glycerophosphate dehydrogenase. Acta Biochim Pol. 2003; 50(2):405-13. View

13.

Lambert A, Buckingham J, Boysen H, Brand M . Diphenyleneiodonium acutely inhibits reactive oxygen species production by mitochondrial complex I during reverse, but not forward electron transport. Biochim Biophys Acta. 2008; 1777(5):397-403. DOI: 10.1016/j.bbabio.2008.03.005. View

14.

Beleznai Z, Jancsik V . Role of cardiolipin in the functioning of mitochondrial L-glycerol-3-phosphate dehydrogenase. Biochem Biophys Res Commun. 1989; 159(1):132-9. DOI: 10.1016/0006-291x(89)92414-5. View

15.

Brown L, MacDonald M, Lehn D, Moran S . Sequence of rat mitochondrial glycerol-3-phosphate dehydrogenase cDNA. Evidence for EF-hand calcium-binding domains. J Biol Chem. 1994; 269(20):14363-6. View

16.

Drahota Z, Saf P, Rauchova H, Kalous M . Dicarbanonaborates--new inhibitors of mitochondrial L-glycerol-3-phosphate dehydrogenase. Biochem Biophys Res Commun. 1995; 210(3):760-5. DOI: 10.1006/bbrc.1995.1724. View

17.

Saheki T, Iijima M, Li M, Kobayashi K, Horiuchi M, Ushikai M . Citrin/mitochondrial glycerol-3-phosphate dehydrogenase double knock-out mice recapitulate features of human citrin deficiency. J Biol Chem. 2007; 282(34):25041-52. DOI: 10.1074/jbc.M702031200. View

18.

Treberg J, Quinlan C, Brand M . Evidence for two sites of superoxide production by mitochondrial NADH-ubiquinone oxidoreductase (complex I). J Biol Chem. 2011; 286(31):27103-10. PMC: 3149303. DOI: 10.1074/jbc.M111.252502. View

19.

Yeh J, Chinte U, Du S . Structure of glycerol-3-phosphate dehydrogenase, an essential monotopic membrane enzyme involved in respiration and metabolism. Proc Natl Acad Sci U S A. 2008; 105(9):3280-5. PMC: 2265192. DOI: 10.1073/pnas.0712331105. View

20.

Maity P, Bindu S, Dey S, Goyal M, Alam A, Pal C . Indomethacin, a non-steroidal anti-inflammatory drug, develops gastropathy by inducing reactive oxygen species-mediated mitochondrial pathology and associated apoptosis in gastric mucosa: a novel role of mitochondrial aconitase oxidation. J Biol Chem. 2008; 284(5):3058-3068. DOI: 10.1074/jbc.M805329200. View