title: Nitric oxide storage and transport in cells are mediated by glutathione S-transferase P1-1 and multidrug resistance protein 1 via dinitrosyl iron complexes.
abstract: Nitrogen monoxide (NO) plays a role in the cytotoxic mechanisms of activated macrophages against tumor cells by inducing iron release. We showed that NO-mediated iron efflux from cells required glutathione (GSH) (Watts, R. N., and Richardson, D. R. (2001) J. Biol. Chem. 276, 4724-4732) and that the GSH-conjugate transporter, multidrug resistance-associated protein 1 (MRP1), mediates this release potentially as a dinitrosyl-dithiol iron complex (DNIC; Watts, R. N., Hawkins, C., Ponka, P., and Richardson, D. R. (2006) Proc. Natl. Acad. Sci. U.S.A. 103, 7670-7675). Recently, glutathione S-transferase P1-1 (GST P1-1) was shown to bind DNICs as dinitrosyl-diglutathionyl iron complexes. Considering this and that GSTs and MRP1 form an integrated detoxification unit with chemotherapeutics, we assessed whether these proteins coordinately regulate storage and transport of DNICs as long lived NO intermediates. Cells transfected with GSTP1 (but not GSTA1 or GSTM1) significantly decreased NO-mediated 59Fe release from cells. This NO-mediated 59Fe efflux and the effect of GST P1-1 on preventing this were observed with NO-generating agents and also in cells transfected with inducible nitric oxide synthase. Notably, 59Fe accumulated in cells within GST P1-1-containing fractions, indicating an alteration in intracellular 59Fe distribution. Furthermore, electron paramagnetic resonance studies showed that MCF7-VP cells transfected with GSTP1 contain significantly greater levels of a unique DNIC signal. These investigations indicate that GST P1-1 acts to sequester NO as DNICs, reducing their transport out of the cell by MRP1. Cell proliferation studies demonstrated the importance of the combined effect of GST P1-1 and MRP1 in protecting cells from the cytotoxic effects of NO. Thus, the DNIC storage function of GST P1-1 and ability of MRP1 to efflux DNICs are vital in protection against NO cytotoxicity.
authors.affiliation: Department of Pathology, University of Sydney, Sydney, New South Wales 2006, Australia.
authors.forename: Hiu Chuen
authors.initials: HC
authors.lastname: Lok
authors.affiliation: Department of Pathology, University of Sydney, Sydney, New South Wales 2006, Australia.
authors.forename: Yohan Suryo
authors.initials: YS
authors.lastname: Rahmanto
authors.affiliation: Heart Research Institute, Sydney, New South Wales 2042, Australia.
authors.forename: Clare L
authors.initials: CL
authors.lastname: Hawkins
authors.affiliation: Department of Pathology, University of Sydney, Sydney, New South Wales 2006, Australia.
authors.forename: Danuta S
authors.initials: DS
authors.lastname: Kalinowski
authors.affiliation: Department of Biochemistry, Wake Forest University, Winston-Salem, North Carolina 27157.
authors.forename: Charles S
authors.initials: CS
authors.lastname: Morrow
authors.affiliation: Department of Biochemistry, Wake Forest University, Winston-Salem, North Carolina 27157.
authors.forename: Alan J
authors.initials: AJ
authors.lastname: Townsend
authors.affiliation: Lady Davis Institute, Montreal, Quebec H3T 1E2, Canada.
authors.forename: Prem
authors.initials: P
authors.lastname: Ponka
authors.affiliation: Department of Pathology, University of Sydney, Sydney, New South Wales 2006, Australia. Electronic address: d.richardson@med.usyd.edu.au.
authors.forename: Des R
authors.initials: DR
authors.lastname: Richardson
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation / Q000187:drug effects; D055785:Gene Knockdown Techniques; D051549:Glutathione S-Transferase pi / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism; D007501:Iron / Q000378:metabolism*; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D009569:Nitric Oxide / Q000378:metabolism* / Q000494:pharmacology; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D009589:Nitrogen Oxides / Q000378:metabolism*; D011422:Propionates / Q000494:pharmacology; D011804:Quinolines / Q000494:pharmacology; D026422:S-Nitrosoglutathione / Q000378:metabolism; D014162:Transfection
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D009589:Nitrogen Oxides; D011422:Propionates; D011804:Quinolines; D009569:Nitric Oxide; D026422:S-Nitrosoglutathione; C059141:verlukast; C073661:dinitrosyl iron complex; D007501:Iron; D052247:Nitric Oxide Synthase Type II; D051549:Glutathione S-Transferase pi
references.citation: Biochem Pharmacol. 1998 Oct 15;56(8):1013-21
references.pmid: 9776312
references.citation: Biochemistry. 2000 Mar 14;39(10):2748-58
references.pmid: 10704227
references.citation: Proc Natl Acad Sci U S A. 2009 Mar 24;106(12):4671-6
references.pmid: 19261856
references.citation: J Neuroimmune Pharmacol. 2010 Jun;5(2):240-51
references.pmid: 20091246
references.citation: Pharmacol Rev. 2005 Dec;57(4):547-83
references.pmid: 16382108
references.citation: J Invest Dermatol. 2003 Sep;121(3):465-74
references.pmid: 12925201
references.citation: J Am Chem Soc. 2008 Nov 19;130(46):15602-10
references.pmid: 18939795
references.citation: J Biol Chem. 2003 Oct 24;278(43):42294-9
references.pmid: 12871931
references.citation: J Biol Chem. 2005 Dec 23;280(51):42172-80
references.pmid: 16195232
references.citation: Biochem J. 2004 May 15;380(Pt 1):67-74
references.pmid: 14766015
references.citation: Biochem J. 2011 Dec 1;440(2):175-83
references.pmid: 21834791
references.citation: Science. 1992 Dec 18;258(5090):1898-902
references.pmid: 1281928
references.citation: Carcinogenesis. 1998 Jan;19(1):109-15
references.pmid: 9472701
references.citation: J Biol Chem. 1979 Aug 25;254(16):7558-60
references.pmid: 38242
references.citation: J Clin Invest. 1986 Sep;78(3):790-7
references.pmid: 3745439
references.citation: Science. 1983 Aug 19;221(4612):769-70
references.pmid: 6308761
references.citation: J Clin Invest. 1999 Feb;103(3):393-9
references.pmid: 9927500
references.citation: Proc Natl Acad Sci U S A. 2001 Jan 2;98(1):355-60
references.pmid: 11134509
references.citation: Int J Cancer. 2001 Jun 15;92(6):777-83
references.pmid: 11351295
references.citation: Mol Pharmacol. 2004 Apr;65(4):897-905
references.pmid: 15044619
references.citation: J Biol Chem. 2007 Mar 2;282(9):6364-71
references.pmid: 17197702
references.citation: Biochem Biophys Res Commun. 1984 Sep 17;123(2):716-23
references.pmid: 6487309
references.citation: Carcinogenesis. 2009 Sep;30(9):1571-80
references.pmid: 19608619
references.citation: Ann Clin Lab Sci. 2002 Summer;32(3):257-63
references.pmid: 12175088
references.citation: J Biol Chem. 2001 Feb 16;276(7):4724-32
references.pmid: 11078730
references.citation: J Biol Chem. 1994 Feb 18;269(7):5127-31
references.pmid: 7508933
references.citation: Biochem J. 2003 Feb 1;369(Pt 3):429-40
references.pmid: 12423201
references.citation: Hypertens Res. 2008 May;31(5):1015-9
references.pmid: 18712057
references.citation: J Biol Chem. 1998 Aug 7;273(32):20114-20
references.pmid: 9685354
references.citation: Br J Cancer. 2005 Jul 25;93(2):216-23
references.pmid: 15999103
references.citation: Free Radic Biol Med. 2011 Oct 15;51(8):1558-66
references.pmid: 21787861
references.citation: Proc Natl Acad Sci U S A. 2006 May 16;103(20):7670-5
references.pmid: 16679408
references.citation: Mol Pharmacol. 2008 Mar;73(3):833-44
references.pmid: 18029550
references.citation: J Biol Chem. 1995 Dec 8;270(49):29244-9
references.pmid: 7493954
references.citation: Biochim Biophys Acta. 2008 Apr;1780(4):638-51
references.pmid: 18206118
references.citation: FEBS Lett. 1991 Sep 2;289(1):1-3
references.pmid: 1893995
references.citation: Proc Natl Acad Sci U S A. 1990 Feb;87(3):1223-7
references.pmid: 2153975