TY - JOUR
T1 - Mutant and wild-type α-synuclein interact with mitochondrial cytochrome C oxidase
AU - Elkon, Hanock
AU - Don, Jermy
AU - Melamed, Eldad
AU - Ziv, Ilan
AU - Shirvan, Anat
AU - Offen, Daniel
N1 - Funding Information:
This work was performed in partial fulfillment of the requirements for a PhD degree of Hanoch Elkon, Sackler Faculty of Medicine, Tel-Aviv University, Israel. Supported, in part, by the National Parkinson Foundation, U.S.A.
PY - 2002
Y1 - 2002
N2 - α-synuclein, a presynaptic protein, was found to be the major component in the Lewy bodies (LB) in both inherited and sporadic Parkinson's disease (PD). Furthermore, rare mutations of α-synuclein cause autosomaldominant PD. However, it is unknown how α-synuclein is involved in the pathogenesis of nigral degeneration in PD. In this study, we examine the protein-protein interactions of wild-type and mutant (A53T) α-synuclein with adult human brain cDNA expression library using the yeast two-hybrid technique. We found that both normal and mutant α-synuclein specifically interact with the mitochondrial complex IV enzyme, cytochrome C oxidase (COX). Wild-type and mutant α-synuclein genes were further fused with c-Myc tag and translated in rabbit reticulocyte lysate. Using anti-c-Myc antibody, we demonstrated that both wild-type and mutant α-synuclein, coimmunoprecipitated with COX. We also showed that potassium cyanide, a selective COX inhibitor, synergistically enhanced the sensitivity of SH-SY5Y neuroblastoma cells to dopamine-induced cell death. In conclusion, we found specific protein-protein interactions of α-synuclein, a major LB protein, to COX, a key enzyme of the mithochondrial respiratory system. This interaction suggests that α-synuclein aggregation may contribute to enhance the mitochondrial dysfunction, which might be a key factor in the pathogenesis of PD.
AB - α-synuclein, a presynaptic protein, was found to be the major component in the Lewy bodies (LB) in both inherited and sporadic Parkinson's disease (PD). Furthermore, rare mutations of α-synuclein cause autosomaldominant PD. However, it is unknown how α-synuclein is involved in the pathogenesis of nigral degeneration in PD. In this study, we examine the protein-protein interactions of wild-type and mutant (A53T) α-synuclein with adult human brain cDNA expression library using the yeast two-hybrid technique. We found that both normal and mutant α-synuclein specifically interact with the mitochondrial complex IV enzyme, cytochrome C oxidase (COX). Wild-type and mutant α-synuclein genes were further fused with c-Myc tag and translated in rabbit reticulocyte lysate. Using anti-c-Myc antibody, we demonstrated that both wild-type and mutant α-synuclein, coimmunoprecipitated with COX. We also showed that potassium cyanide, a selective COX inhibitor, synergistically enhanced the sensitivity of SH-SY5Y neuroblastoma cells to dopamine-induced cell death. In conclusion, we found specific protein-protein interactions of α-synuclein, a major LB protein, to COX, a key enzyme of the mithochondrial respiratory system. This interaction suggests that α-synuclein aggregation may contribute to enhance the mitochondrial dysfunction, which might be a key factor in the pathogenesis of PD.
KW - Coimmunoprecipitation
KW - Cytochrome c oxidase
KW - In vitro translation
KW - Parkinson's disease
KW - Potassium cyanide
KW - Two-hybrid system
KW - α-synuclein
UR - http://www.scopus.com/inward/record.url?scp=0036014117&partnerID=8YFLogxK
U2 - 10.1385/JMN:18:3:229
DO - 10.1385/JMN:18:3:229
M3 - ???researchoutput.researchoutputtypes.contributiontojournal.article???
AN - SCOPUS:0036014117
SN - 0895-8696
VL - 18
SP - 229
EP - 238
JO - Journal of Molecular Neuroscience
JF - Journal of Molecular Neuroscience
IS - 3
ER -