title: Identification of proteins adducted by lipid peroxidation products in plasma and modifications of apolipoprotein A1 with a novel biotinylated phospholipid probe.
abstract: Reactive electrophiles generated by lipid peroxidation are thought to contribute to cardiovascular disease and other oxidative stress-related pathologies by covalently modifying proteins and affecting critical protein functions. The difficulty of capturing and analyzing the relatively small fraction of modified proteins complicates identification of the protein targets of lipid electrophiles. We recently synthesized a biotin-modified linoleoylglycerylphosphatidylcholine probe called PLPBSO ( Tallman et al. Chem. Res. Toxicol. 2007, 20, 227-234 ), which forms typical linoleate oxidation products and covalent adducts with model peptides and proteins. Supplementation of human plasma with PLPBSO followed by free radical oxidation resulted in covalent adduction of PLPBSO to plasma proteins, which were isolated with streptavidin and identified by liquid chromatography-tandem mass spectrometry (LC-MS-MS). Among the most highly modified proteins was apolipoprotein A1 (ApoA1), which is the core component of high density lipoprotein (HDL). ApoA1 phospholipid adduct sites were mapped by LC-MS-MS of tryptic peptides following mild base hydrolysis to release esterified phospholipid adducts. Several carboxylated adducts formed from phospholipid-esterified 9,12-dioxo-10( E)-dodecenoic acid (KODA), 9-hydroxy, 12-oxo-10( E)-dodecenoic acid (HODA), 7-oxoheptanoic acid, 8-oxooctanoic acid, and 9-oxononanoic acid were identified. Free radical oxidations of isolated HDL also generated adducts with 4-hydroxynonenal (HNE) and other noncarboxylated electrophiles, but these were only sporadically identified in the PLPBSO-adducted ApoA1, suggesting a low stoichiometry of modification in the phospholipid-adducted protein. Both phospholipid electrophiles and HNE adducted His162, which resides in an ApoA1 domain involved in the activation of Lecithin-cholesterol acyltransferase and maturation of the HDL particle. ApoA1 lipid electrophile adducts may affect protein functions and provide useful biomarkers for oxidative stress.
authors.affiliation: Department of Biochemistry, Vanderbilt University, Nashville, Tennessee 37232-0146, USA.
authors.forename: Matthew E
authors.initials: ME
authors.lastname: Szapacs
authors.forename: Hye-Young H
authors.initials: HY
authors.lastname: Kim
authors.forename: Ned A
authors.initials: NA
authors.lastname: Porter
authors.forename: Daniel C
authors.initials: DC
authors.lastname: Liebler
mesh_terms: D016632:Apolipoprotein A-I / Q000737:chemistry* / Q000378:metabolism; D001710:Biotin / Q000737:chemistry / Q000378:metabolism*; D001798:Blood Proteins / Q000737:chemistry / Q000378:metabolism; D002853:Chromatography, Liquid; D006801:Humans; D015227:Lipid Peroxidation; D013058:Mass Spectrometry; D015335:Molecular Probes / Q000737:chemistry / Q000378:metabolism; D015394:Molecular Structure; D010713:Phosphatidylcholines / Q000737:chemistry* / Q000378:metabolism*; D010743:Phospholipids / Q000737:chemistry / Q000378:metabolism; D010949:Plasma / Q000737:chemistry*
chemical_list: D016632:Apolipoprotein A-I; D001798:Blood Proteins; D015335:Molecular Probes; D010713:Phosphatidylcholines; D010743:Phospholipids; C532858:biotinylated linoleoylglycerylphosphatidylcholine; D001710:Biotin
references.citation: J Biol Chem. 2002 Oct 11;277(41):38503-16
references.pmid: 12105195
references.citation: J Biol Chem. 2005 Feb 18;280(7):5983-93
references.pmid: 15574409
references.citation: J Pharmacol Exp Ther. 2005 Oct;315(1):8-15
references.pmid: 15951401
references.citation: J Am Soc Mass Spectrom. 1994 Nov;5(11):976-89
references.pmid: 24226387
references.citation: J Clin Invest. 2003 Mar;111(5):583-93
references.pmid: 12618510
references.citation: Chem Res Toxicol. 2007 Feb;20(2):165-70
references.pmid: 17305402
references.citation: Curr Opin Lipidol. 2006 Aug;17(4):399-403
references.pmid: 16832163
references.citation: Chem Res Toxicol. 2005 Aug;18(8):1219-31
references.pmid: 16097795
references.citation: Science. 2001 Jun 15;292(5524):2083-6
references.pmid: 11408659
references.citation: FASEB J. 1999 Jul;13(10):1157-68
references.pmid: 10385607
references.citation: Chem Res Toxicol. 2007 Feb;20(2):227-34
references.pmid: 17305406
references.citation: J Biol Chem. 2006 Apr 7;281(14):9001-4
references.pmid: 16497665
references.citation: J Biol Chem. 2004 Apr 2;279(14):13447-51
references.pmid: 14715668
references.citation: J Biol Chem. 2002 Oct 11;277(41):38517-23
references.pmid: 12145296
references.citation: Lancet. 1994 Sep 10;344(8924):721-4
references.pmid: 7915779
references.citation: J Proteome Res. 2005 Mar-Apr;4(2):358-68
references.pmid: 15822911
references.citation: Circulation. 2004 Jun 15;109(23 Suppl 1):III8-14
references.pmid: 15198960
references.citation: Chem Res Toxicol. 2007 Jun;20(6):849-53
references.pmid: 17489606
references.citation: Mutat Res. 2001 Apr 18;475(1-2):7-20
references.pmid: 11295149
references.citation: J Biol Chem. 2002 Mar 1;277(9):7010-20
references.pmid: 11744722
references.citation: J Biol Chem. 1999 Aug 27;274(35):24787-98
references.pmid: 10455151
references.citation: Arterioscler Thromb Vasc Biol. 2000 Oct;20(10):2248-54
references.pmid: 11031211
references.citation: J Biol Chem. 1997 May 23;272(21):13597-607
references.pmid: 9153208
references.citation: J Biol Chem. 2005 Oct 28;280(43):36386-96
references.pmid: 16126721
references.citation: Pharmacol Rev. 2006 Sep;58(3):342-74
references.pmid: 16968945
references.citation: Chem Res Toxicol. 2007 Jan;20(1):3-5
references.pmid: 17226920
references.citation: Chem Res Toxicol. 2007 May;20(5):767-75
references.pmid: 17407328
references.citation: Physiol Rev. 1998 Apr;78(2):547-81
references.pmid: 9562038
references.citation: Free Radic Biol Med. 2002 Dec 1;33(11):1475-9
references.pmid: 12446204
references.citation: Lipids. 1995 Apr;30(4):277-90
references.pmid: 7609594
references.citation: Chem Res Toxicol. 2003 Jul;16(7):901-11
references.pmid: 12870893
references.citation: J Biol Chem. 1996 Nov 8;271(45):27999-8001
references.pmid: 8910407
references.citation: J Biol Chem. 1999 May 7;274(19):13139-46
references.pmid: 10224068
references.citation: Proteomics. 2007 Feb;7(4):628-640
references.pmid: 17309108
references.citation: Methods Enzymol. 1986;128:155-70
references.pmid: 3724500
references.citation: Proteomics. 2001 Jan;1(1):93-107
references.pmid: 11680902
references.citation: Trends Cardiovasc Med. 2005 Aug;15(6):212-9
references.pmid: 16182131
references.citation: N Engl J Med. 2005 Jul 7;353(1):9-11
references.pmid: 16000351
references.citation: Biochemistry. 2003 Apr 22;42(15):4398-405
references.pmid: 12693935
references.citation: Mol Pharmacol. 2007 Mar;71(3):871-83
references.pmid: 17164404
references.citation: J Clin Invest. 2004 Aug;114(4):529-41
references.pmid: 15314690
references.citation: Science. 1983 Sep 23;221(4617):1256-64
references.pmid: 6351251
references.citation: Free Radic Biol Med. 2005 Mar 15;38(6):698-710
references.pmid: 15721980
references.citation: Proc Natl Acad Sci U S A. 1999 May 25;96(11):6353-8
references.pmid: 10339591
references.citation: J Biol Chem. 2002 Mar 1;277(9):7271-81
references.pmid: 11751881
references.citation: Chem Res Toxicol. 2004 Nov;17(11):1459-67
references.pmid: 15540944
references.citation: Nat Med. 2007 Sep;13(9):1086-95
references.pmid: 17721545
references.citation: Chem Res Toxicol. 2005 Aug;18(8):1324-31
references.pmid: 16097806
references.citation: J Biol Chem. 2005 Oct 7;280(40):33775-84
references.pmid: 16091367
references.citation: Biochemistry. 2006 Sep 5;45(35):10521-8
references.pmid: 16939204
references.citation: Ann N Y Acad Sci. 2005 Jun;1043:396-403
references.pmid: 16037261
references.citation: Nat Struct Mol Biol. 2007 Sep;14(9):861-8
references.pmid: 17676061
references.citation: Arterioscler Thromb Vasc Biol. 2005 Jun;25(6):1102-11
references.pmid: 15790935
references.citation: Biochemistry. 1999 Oct 26;38(43):14409-15
references.pmid: 10572015
references.citation: Free Radic Res. 2002 Dec;36(12):1307-13
references.pmid: 12607822
references.citation: J Am Soc Mass Spectrom. 1996 Jun;7(6):532-40
references.pmid: 24203425
references.citation: J Biol Chem. 2005 Jan 7;280(1):38-47
references.pmid: 15498770
references.citation: Chem Res Toxicol. 2002 Nov;15(11):1445-50
references.pmid: 12437335
references.citation: Proc Natl Acad Sci U S A. 2002 Oct 1;99(20):13043-8
references.pmid: 12244213
references.citation: J Biol Chem. 2001 Jun 15;276(24):20831-8
references.pmid: 11259420