Mutant and wild-type alpha-synuclein interact with mitochondrial cytochrome C oxidase

J Mol Neurosci. 2002 Jun;18(3):229-38. doi: 10.1385/JMN:18:3:229.

Abstract

Alpha-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 alpha-synuclein cause autosomal-dominant PD. However, it is unknown how alpha-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) a-synuclein with adult human brain cDNA expression library using the yeast two-hybrid technique. We found that both normal and mutant alpha-synuclein specifically interact with the mitochondrial complex IV enzyme, cytochrome C oxidase (COX). Wild-type and mutant alpha-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 alpha-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 alpha-synuclein, a major LB protein, to COX, a key enzyme of the mithochondrial respiratory system. This interaction suggests that alpha-synuclein aggregation may contribute to enhance the mitochondrial dysfunction, which might be a key factor in the pathogenesis of PD.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Base Sequence
  • Dopamine / metabolism
  • Electron Transport Complex IV / antagonists & inhibitors
  • Electron Transport Complex IV / metabolism*
  • Enzyme Inhibitors / metabolism
  • Humans
  • Mitochondria / enzymology*
  • Molecular Sequence Data
  • Mutation
  • Nerve Tissue Proteins / genetics*
  • Nerve Tissue Proteins / metabolism*
  • Parkinson Disease / metabolism*
  • Potassium Cyanide / metabolism
  • Protein Binding
  • Proto-Oncogene Proteins c-myc / genetics
  • Proto-Oncogene Proteins c-myc / metabolism
  • Synucleins
  • Tumor Cells, Cultured
  • Two-Hybrid System Techniques
  • alpha-Synuclein

Substances

  • Enzyme Inhibitors
  • Nerve Tissue Proteins
  • Proto-Oncogene Proteins c-myc
  • SNCA protein, human
  • Synucleins
  • alpha-Synuclein
  • Electron Transport Complex IV
  • Potassium Cyanide
  • Dopamine