The DJ-1L166P mutant protein associated with early onset Parkinson's disease is unstable and forms higher-order protein complexes

Hum Mol Genet. 2003 Nov 1;12(21):2807-16. doi: 10.1093/hmg/ddg304. Epub 2003 Sep 2.

Abstract

Parkinson's disease (PD) is a common neurodegenerative disorder that involves the selective degeneration of midbrain dopaminergic neurons. Recently DJ-1 mutations have been linked to autosomal-recessive early-onset Parkinsonism in two European families. By using gel filtration assays under physiological conditions we demonstrate that DJ-1 protein forms a dimeric structure. Conversely, the DJ-1L166P mutant protein shows a different elution profile as compared with DJ-1WT both in overexpression cellular systems or in lymphoblasts cells, suggesting that it might form higher order protein structures. Furthermore we observed that the level of DJ-1L166P mutant protein in the patient's lymphoblasts was very low as compared with the wild-type protein. We excluded a potential transcriptional impairment by performing quantitative RT-PCR on the patient's material. Pulse-chase experiments in transfected COS-1 cells and cycloheximide treatment in control and patient lymphoblasts indicated that the mutant protein was rapidly degraded. This rapid turnover and the structural changes of DJ-1L166P mutant protein might be crucial in the disease pathogenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • COS Cells
  • Chlorocebus aethiops
  • Cycloheximide / pharmacology
  • Dimerization
  • HeLa Cells
  • Humans
  • Intracellular Signaling Peptides and Proteins
  • Lymphocytes / metabolism*
  • Mutation
  • Nerve Tissue Proteins / metabolism
  • Nuclear Proteins / metabolism
  • Oncogene Proteins / genetics*
  • Oncogene Proteins / metabolism
  • Parkinson Disease / genetics*
  • Parkinson Disease / metabolism
  • Protein Deglycase DJ-1
  • Protein Denaturation
  • Protein Inhibitors of Activated STAT
  • Protein Synthesis Inhibitors / pharmacology
  • Synucleins

Substances

  • Intracellular Signaling Peptides and Proteins
  • Nerve Tissue Proteins
  • Nuclear Proteins
  • Oncogene Proteins
  • PIAS2 protein, human
  • Protein Inhibitors of Activated STAT
  • Protein Synthesis Inhibitors
  • Synucleins
  • Cycloheximide
  • PARK7 protein, human
  • Protein Deglycase DJ-1