title: Reduction of mitochondrial protein mitoNEET [2Fe-2S] clusters by human glutathione reductase.
abstract: The human mitochondrial outer membrane protein mitoNEET is a newly discovered target of the type 2 diabetes drug pioglitazone. Structurally, mitoNEET is a homodimer with each monomer containing an N-terminal transmembrane α helix tethered to the mitochondrial outer membrane and a C-terminal cytosolic domain hosting a redox-active [2Fe-2S] cluster. Genetic studies have shown that mitoNEET has a central role in regulating energy metabolism in mitochondria. However, the specific function of mitoNEET remains largely elusive. Here we find that the mitoNEET [2Fe-2S] clusters can be efficiently reduced by Escherichia coli thioredoxin reductase and glutathione reductase in an NADPH-dependent reaction. Purified human glutathione reductase has the same activity as E. coli thioredoxin reductase and glutathione reductase to reduce the mitoNEET [2Fe-2S] clusters. However, rat thioredoxin reductase, a human thioredoxin reductase homolog that contains selenocysteine in the catalytic center, has very little or no activity to reduce the mitoNEET [2Fe-2S] clusters. N-ethylmaleimide, a potent thiol modifier, completely inhibits human glutathione reductase from reducing the mitoNEET [2Fe-2S] clusters, indicating that the redox-active disulfide in the catalytic center of human glutathione reductase may be directly involved in reducing the mitoNEET [2Fe-2S] clusters. Additional studies reveal that the reduced mitoNEET [2Fe-2S] clusters in mouse heart cell extracts can be reversibly oxidized by hydrogen peroxide without disruption of the clusters, suggesting that the mitoNEET [2Fe-2S] clusters may undergo redox transition to regulate energy metabolism in mitochondria in response to oxidative signals.
authors: Department of Biological Sciences, Louisiana State University, Baton Rouge, LA 70803, USA.
authors: Aaron P
authors: AP
authors: Landry
authors: Department of Biological Sciences, Louisiana State University, Baton Rouge, LA 70803, USA.
authors: Zishuo
authors: Z
authors: Cheng
authors: Department of Biological Sciences, Louisiana State University, Baton Rouge, LA 70803, USA. Electronic address: hding@lsu.edu.
authors: Huangen
authors: H
authors: Ding
mesh_terms: D000818:Animals; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D005033:Ethylmaleimide / Q000494:pharmacology; D015870:Gene Expression; D005980:Glutathione Reductase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007506:Iron-Sulfur Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008929:Mitochondria, Heart / Q000187:drug effects / Q000378:metabolism*; D051336:Mitochondrial Membranes / Q000187:drug effects / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009206:Myocardium / Q000737:chemistry; D009249:NADP / Q000737:chemistry / Q000378:metabolism; D010084:Oxidation-Reduction; D000077205:Pioglitazone; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D045162:Thiazolidinediones / Q000494:pharmacology; D013880:Thioredoxin-Disulfide Reductase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: C525595:CISD1 protein, human; D029968:Escherichia coli Proteins; D007506:Iron-Sulfur Proteins; D024101:Mitochondrial Proteins; D011994:Recombinant Proteins; D045162:Thiazolidinediones; D009249:NADP; D006861:Hydrogen Peroxide; D005980:Glutathione Reductase; D013880:Thioredoxin-Disulfide Reductase; D005033:Ethylmaleimide; D000077205:Pioglitazone
keywords: Free radicals; Glutathione reductase; Iron–sulfur cluster; MitoNEET; Thioredoxin reductase; Type 2 diabetes
references: Parasitol Int. 2013 Apr;62(2):193-8
references: 23305756
references: Mol Endocrinol. 2014 Nov;28(11):1756-68
references: 25148456
references: Drug Discov Today. 2014 Oct;19(10):1601-6
references: 24814435
references: Eur J Biochem. 2000 Oct;267(20):6102-9
references: 11012661
references: Int J Obes Relat Metab Disord. 2003 Feb;27(2):147-61
references: 12586994
references: Am J Physiol Endocrinol Metab. 2004 Feb;286(2):E252-60
references: 14570702
references: J Biol Chem. 2004 May 21;279(21):22284-93
references: 14985369
references: Anal Biochem. 1976 May 7;72:248-54
references: 942051
references: Anal Biochem. 1980 Jul 15;106(1):207-12
references: 7416462
references: J Biol Chem. 2014 Oct 10;289(41):28070-86
references: 25012650
references: Biochemistry. 2009 Nov 3;48(43):10193-5
references: 19791753
references: Proc Natl Acad Sci U S A. 2000 May 23;97(11):5854-9
references: 10801974
references: J Biol Chem. 2000 Jun 16;275(24):18121-8
references: 10849437
references: PLoS One. 2010;5(2):e9280
references: 20174634
references: Proc Natl Acad Sci U S A. 2011 Mar 29;108(13):5266-71
references: 21402934
references: Biochemistry. 2010 Nov 9;49(44):9604-12
references: 20932062
references: Nat Commun. 2011;2:383
references: 21750537
references: Proc Natl Acad Sci U S A. 2011 Aug 9;108(32):13047-52
references: 21788481
references: PLoS One. 2011;6(9):e24790
references: 21949752
references: Proc Natl Acad Sci U S A. 2012 Feb 7;109(6):1955-60
references: 22308404
references: J Biol Chem. 2012 Apr 6;287(15):11649-55
references: 22351774
references: Plant Cell. 2012 May;24(5):2139-54
references: 22562611
references: Cell. 2012 Aug 31;150(5):1068-81
references: 22939629
references: Nat Med. 2012 Oct;18(10):1539-49
references: 22961109
references: Biometals. 2012 Dec;25(6):1177-84
references: 22945239
references: Biochim Biophys Acta. 2013 May;1830(5):3217-66
references: 23036594
references: Nature. 2013 Apr 18;496(7445):372-6
references: 23503661
references: J Biol Chem. 2013 Apr 19;288(16):11304-11
references: 23479729
references: EMBO Mol Med. 2013 Jun;5(6):904-18
references: 23703906
references: Proc Natl Acad Sci U S A. 2013 Sep 3;110(36):14676-81
references: 23959881
references: Biochemistry. 2013 Jul 9;52(27):4687-96
references: 23758282
references: Biochemistry. 2013 Dec 17;52(50):8969-71
references: 24295216
references: Biochemistry. 2014 Jan 28;53(3):554-65
references: 24393022
references: J Biol Chem. 2014 Feb 14;289(7):4307-15
references: 24403080
references: J Cell Sci. 2014 Feb 15;127(Pt 4):896-907
references: 24357718
references: J Biol Chem. 2014 Feb 21;289(8):5145-57
references: 24379410
references: Proc Biol Sci. 1990 Sep 22;241(1302):179-86
references: 1979442
references: Arch Biochem Biophys. 1992 May 1;294(2):403-6
references: 1314540
references: FASEB J. 1995 Oct;9(13):1267-76
references: 7557016
references: Protein Expr Purif. 1997 Apr;9(3):372-8
references: 9126609
references: J Mol Biol. 1999 Oct 8;292(5):1003-16
references: 10512699
references: J Biol Chem. 2005 May 27;280(21):20628-37
references: 15792952
references: Biochem Pharmacol. 2005 Jul 15;70(2):177-88
references: 15925327
references: Annu Rev Pharmacol Toxicol. 2006;46:215-34
references: 16402904
references: DNA Res. 2005;12(5):291-9
references: 16769691
references: J Biol Chem. 2006 Aug 11;281(32):22953-63
references: 16774913
references: Proc Natl Acad Sci U S A. 2007 Mar 27;104(13):5318-23
references: 17376863
references: J Biol Chem. 2007 Aug 17;282(33):23745-9
references: 17584744
references: Proc Natl Acad Sci U S A. 2007 Sep 4;104(36):14342-7
references: 17766440
references: Am J Hum Genet. 2007 Oct;81(4):673-83
references: 17846994
references: Proc Natl Acad Sci U S A. 2007 Sep 11;104(37):14640-5
references: 17766439
references: J Biol Chem. 2007 Nov 16;282(46):33242-6
references: 17905743
references: Biochemistry. 2008 Jan 29;47(4):1167-75
references: 18171025
references: Annu Rev Biochem. 2008;77:289-312
references: 18518822
references: J Proteome Res. 2008 Oct;7(10):4566-76
references: 18781797
references: Acta Crystallogr Sect F Struct Biol Cryst Commun. 2008 Dec 1;64(Pt 12):1146-8
references: 19052371
references: Biochemistry. 2009 Feb 10;48(5):1016-24
references: 19132843
references: Genes Dev. 2009 May 15;23(10):1183-94
references: 19451219
references: Biochemistry. 2009 Jun 9;48(22):4747-52
references: 19388667
references: Biochim Biophys Acta. 2009 Jun;1790(6):495-526
references: 19364476
references: Nature. 2009 Aug 13;460(7257):831-8
references: 19675643
references: J Mol Biol. 2009 Sep 11;392(1):143-53
references: 19580816
references: Nat Commun. 2014;5:3962
references: 24865177