Preferentially increased nitration of alpha-synuclein at tyrosine-39 in a cellular oxidative model of Parkinson's disease.
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.
Danielson
Steven R
SR
Buck Institute for Age Research, 8001 Redwood Boulevard, Novato, California 94945, USA.
Held
Jason M
JM
Schilling
Birgit
B
Oo
May
M
Gibson
Bradford W
BW
Andersen
Julie K
JK
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
D009566:Nitrates; D051844:alpha-Synuclein; C002744:3-nitrotyrosine; D014443:Tyrosine; D008995:Monoamine Oxidase
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