Abstract
Parkinson's disease is a movement disorder that results from a loss of dopaminergic neurons in the substantia nigra. The disease is characterized by mitochondrial dysfunction, oxidative stress, and the presence of "Lewy body" inclusions enriched with aggregated forms of α-synuclein, a presynaptic protein. Although α-synuclein is modified at various sites in Lewy bodies, it is unclear how sequence-specific posttranslational modifications modulate the aggregation of the protein in oxidatively stressed neurons. To begin to address this problem, we developed an affinity pull-down/mass spectrometry method to characterize the primary structure of histidine-tagged α-synuclein isolated from catecholaminergic neurons. Using this method, we mapped posttranslational modifications of α-synuclein from untreated neurons and neurons exposed to rotenone, an inhibitor of mitochondrial complex I. Various posttranslational modifications suggestive of oxidative damage or repair were identified in a region comprising a 20-residue stretch in the C-terminal part of the protein. The results indicate that α-synuclein is subject to discrete posttranslational modifications in neurons with impaired mitochondrial function. Our affinity pull-down/mass spectrometry method is a useful tool to examine how specific modifications of α-synuclein contribute to neurologic disorders such as Parkinson's disease.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 2422-2431 |
| Number of pages | 10 |
| Journal | Analytical Chemistry |
| Volume | 78 |
| Issue number | 7 |
| DOIs | |
| State | Published - Apr 1 2006 |
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- Analytical Chemistry
Cite this
Identification of rotenone-induced modifications in α-synuclein using affinity pull-down and tandem mass spectrometry. / Mirzaei, Hamid; Schieler, Jeremy L.; Rochet, Jean Christophe; Regnier, Fred.
In: Analytical Chemistry, Vol. 78, No. 7, 01.04.2006, p. 2422-2431.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Identification of rotenone-induced modifications in α-synuclein using affinity pull-down and tandem mass spectrometry
AU - Mirzaei, Hamid
AU - Schieler, Jeremy L.
AU - Rochet, Jean Christophe
AU - Regnier, Fred
PY - 2006/4/1
Y1 - 2006/4/1
N2 - Parkinson's disease is a movement disorder that results from a loss of dopaminergic neurons in the substantia nigra. The disease is characterized by mitochondrial dysfunction, oxidative stress, and the presence of "Lewy body" inclusions enriched with aggregated forms of α-synuclein, a presynaptic protein. Although α-synuclein is modified at various sites in Lewy bodies, it is unclear how sequence-specific posttranslational modifications modulate the aggregation of the protein in oxidatively stressed neurons. To begin to address this problem, we developed an affinity pull-down/mass spectrometry method to characterize the primary structure of histidine-tagged α-synuclein isolated from catecholaminergic neurons. Using this method, we mapped posttranslational modifications of α-synuclein from untreated neurons and neurons exposed to rotenone, an inhibitor of mitochondrial complex I. Various posttranslational modifications suggestive of oxidative damage or repair were identified in a region comprising a 20-residue stretch in the C-terminal part of the protein. The results indicate that α-synuclein is subject to discrete posttranslational modifications in neurons with impaired mitochondrial function. Our affinity pull-down/mass spectrometry method is a useful tool to examine how specific modifications of α-synuclein contribute to neurologic disorders such as Parkinson's disease.
AB - Parkinson's disease is a movement disorder that results from a loss of dopaminergic neurons in the substantia nigra. The disease is characterized by mitochondrial dysfunction, oxidative stress, and the presence of "Lewy body" inclusions enriched with aggregated forms of α-synuclein, a presynaptic protein. Although α-synuclein is modified at various sites in Lewy bodies, it is unclear how sequence-specific posttranslational modifications modulate the aggregation of the protein in oxidatively stressed neurons. To begin to address this problem, we developed an affinity pull-down/mass spectrometry method to characterize the primary structure of histidine-tagged α-synuclein isolated from catecholaminergic neurons. Using this method, we mapped posttranslational modifications of α-synuclein from untreated neurons and neurons exposed to rotenone, an inhibitor of mitochondrial complex I. Various posttranslational modifications suggestive of oxidative damage or repair were identified in a region comprising a 20-residue stretch in the C-terminal part of the protein. The results indicate that α-synuclein is subject to discrete posttranslational modifications in neurons with impaired mitochondrial function. Our affinity pull-down/mass spectrometry method is a useful tool to examine how specific modifications of α-synuclein contribute to neurologic disorders such as Parkinson's disease.
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UR - http://www.scopus.com/inward/citedby.url?scp=33645680940&partnerID=8YFLogxK
U2 - 10.1021/ac051978n
DO - 10.1021/ac051978n
M3 - Article
C2 - 16579629
AN - SCOPUS:33645680940
VL - 78
SP - 2422
EP - 2431
JO - Analytical Chemistry
JF - Analytical Chemistry
SN - 0003-2700
IS - 7
ER -