title: Preferentially increased nitration of alpha-synuclein at tyrosine-39 in a cellular oxidative model of Parkinson's disease.
abstract: Alpha-synuclein is a major component of Lewy bodies, proteinacious inclusions which are a major hallmark of Parkinson's disease (PD). Lewy bodies contain high levels of nitrated tyrosine residues as determined by antibodies specific for 3-nitrotyrosine (3NT) and via mass spectrometry (MS). We have developed a multiple reaction monitoring (MRM) mass spectrometry method to sensitively quantitate the 3NT levels of specific alpha-synuclein tyrosine residues. We found a 9-fold increase (relative to controls) in levels of 3NT at Tyr-39 of alpha-synuclein in an inducible transgenic cellular model of Parkinson's disease in which monoamine oxidase B (MAO-B) is overexpressed and which emulates several features of PD. Increased nitration of Tyr-39 on endogenous alpha-synuclein via elevations in MAO-B levels could be abrogated by the addition of deprenyl, a specific MAO-B inhibitor. The increased levels of 3NT was selective for Tyr-39 as no significant increases in 3NT levels were detected at other tyrosine residues present in the protein (Tyr-125, Tyr-133, and Tyr-136). This is the first report of increased 3NT levels of a specific tyrosine in a PD model and the first use of MRM mass spectrometry to quantify changes in 3NT modifications at specific sites within a target protein.
authors.affiliation: Buck Institute for Age Research, 8001 Redwood Boulevard, Novato, California 94945, USA.
authors.forename: Steven R
authors.initials: SR
authors.lastname: Danielson
authors.forename: Jason M
authors.initials: JM
authors.lastname: Held
authors.forename: Birgit
authors.initials: B
authors.lastname: Schilling
authors.forename: May
authors.initials: M
authors.lastname: Oo
authors.forename: Bradford W
authors.initials: BW
authors.lastname: Gibson
authors.forename: Julie K
authors.initials: JK
authors.lastname: Andersen
mesh_terms: D000818:Animals; D016631:Lewy Bodies / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods*; D008954:Models, Biological; D008995:Monoamine Oxidase / Q000032:analysis; D009566:Nitrates / Q000378:metabolism*; D018384:Oxidative Stress; D016716:PC12 Cells; D010300:Parkinson Disease / Q000378:metabolism*; D051381:Rats; D014443:Tyrosine / Q000031:analogs & derivatives / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D051844:alpha-Synuclein / Q000737:chemistry* / Q000378:metabolism
chemical_list: D009566:Nitrates; D051844:alpha-Synuclein; C002744:3-nitrotyrosine; D014443:Tyrosine; D008995:Monoamine Oxidase
references.citation: Prog Neurobiol. 2005 Jun;76(2):126-52
references.pmid: 16115721
references.citation: Mol Cell Proteomics. 2009 Mar;8(3):467-80
references.pmid: 18984578
references.citation: Biochemistry. 2008 Dec 23;47(51):13604-9
references.pmid: 19049426
references.citation: Clin Neuropharmacol. 1986;9(6):485-507
references.pmid: 3542203
references.citation: Cell. 1994 Jun 17;77(6):817-27
references.pmid: 8004671
references.citation: J Biol Chem. 2004 Nov 12;279(46):47746-53
references.pmid: 15364911
references.citation: Biochem Soc Trans. 2003 Dec;31(Pt 6):1417-22
references.pmid: 14641078
references.citation: Brain Res. 1986 Jun 18;376(1):117-24
references.pmid: 3487376
references.citation: Am J Physiol Heart Circ Physiol. 2008 Dec;295(6):H2417-26
references.pmid: 18931031
references.citation: J Neurochem. 2007 Oct;103(1):17-37
references.pmid: 17623039
references.citation: Science. 1997 Jun 27;276(5321):2045-7
references.pmid: 9197268
references.citation: J Biol Chem. 2000 Jul 14;275(28):21409-15
references.pmid: 10770952
references.citation: Mech Ageing Dev. 1983 Oct;23(2):151-60
references.pmid: 6140331
references.citation: Biochim Biophys Acta. 2006 Feb;1762(2):213-22
references.pmid: 16120479
references.citation: Neurol Res. 2005 Sep;27(6):630-3
references.pmid: 16157014
references.citation: J Neurochem. 2001 Jan;76(2):637-40
references.pmid: 11208927
references.citation: FEBS Lett. 2003 May 8;542(1-3):147-52
references.pmid: 12729915
references.citation: Neuroreport. 1997 May 27;8(8):2053-60
references.pmid: 9223101
references.citation: J Clin Invest. 2003 Jan;111(2):163-9
references.pmid: 12531868
references.citation: Brain. 1965 Mar;88:181-92
references.pmid: 14280273
references.citation: J Biol Chem. 2004 Mar 12;279(11):10128-35
references.pmid: 14699135
references.citation: J Neurochem. 1992 Nov;59(5):1609-23
references.pmid: 1402908
references.citation: J Biol Chem. 2000 Jun 16;275(24):18344-9
references.pmid: 10747881
references.citation: Ann Neurol. 1992 Feb;31(2):119-30
references.pmid: 1349466
references.citation: Brain Res Mol Brain Res. 2005 Mar 24;134(1):84-102
references.pmid: 15790533
references.citation: Electrophoresis. 1999 Dec;20(18):3551-67
references.pmid: 10612281
references.citation: Protein Sci. 2004 Nov;13(11):2852-6
references.pmid: 15459341
references.citation: J Neurochem. 1993 Jun;60(6):2046-58
references.pmid: 8492117
references.citation: Mol Cell Proteomics. 2009 May;8(5):1006-15
references.pmid: 19068476
references.citation: J Neuropathol Exp Neurol. 1998 Apr;57(4):338-42
references.pmid: 9600227
references.citation: Mol Cell Proteomics. 2005 Jan;4(1):84-96
references.pmid: 15591592
references.citation: Arch Biochem Biophys. 2008 Oct 15;478(2):181-6
references.pmid: 18706882
references.citation: Science. 2000 Nov 3;290(5493):985-9
references.pmid: 11062131
references.citation: Klin Wochenschr. 1960 Dec 15;38:1236-9
references.pmid: 13726012
references.citation: Proc Natl Acad Sci U S A. 2007 Apr 3;104(14):5860-5
references.pmid: 17389395
references.citation: J Biol Chem. 2003 Nov 21;278(47):46432-9
references.pmid: 12963742
references.citation: Brain Res. 2002 May 31;938(1-2):73-80
references.pmid: 12031537
references.citation: Lancet. 1983 Dec 24-31;2(8365-66):1457-9
references.pmid: 6140548
references.citation: Mol Neurobiol. 2004 Aug;30(1):77-89
references.pmid: 15247489
references.citation: Ann Neurol. 1995 Jun;37(6):714-22
references.pmid: 7778844
references.citation: Curr Protein Pept Sci. 2008 Oct;9(5):507-40
references.pmid: 18855701
references.citation: Biochemistry. 2006 Jul 4;45(26):8009-22
references.pmid: 16800626
references.citation: Am J Pathol. 1998 Apr;152(4):879-84
references.pmid: 9546347
references.citation: Neuroreport. 1994 Jul 21;5(12):1485-8
references.pmid: 7948844
references.citation: Nat Med. 2005 Nov;11(11):1214-21
references.pmid: 16227987
references.citation: J Pharm Biomed Anal. 2006 Jun 16;41(4):1325-31
references.pmid: 16616826
references.citation: Ann Neurol. 1998 Sep;44(3 Suppl 1):S72-84
references.pmid: 9749577
references.citation: Mol Cell Proteomics. 2008 Dec;7(12):2442-51
references.pmid: 18708664
references.citation: Nat Neurosci. 2000 Dec;3(12):1301-6
references.pmid: 11100151
references.citation: J Biol Chem. 2006 Oct 6;281(40):29739-52
references.pmid: 16847063