Engineered disulfide bonds restore chaperone-like function of DJ-1 mutants linked to familial Parkinson's disease.
Loss-of-function mutations such as L166P, A104T, and M26I in the DJ-1 gene (PARK7) have been linked to autosomal-recessive early onset Parkinson's disease (PD). Cellular and structural studies of the familial mutants suggest that these mutations may destabilize the dimeric structure. To look for common dynamical signatures among the DJ-1 mutants, short MD simulations of up to 1000 ps were conducted to identify the weakest region of the protein (residues 38-70). In an attempt to stabilize the protein, we mutated residue Val 51 to cysteine (V51C) to make a symmetry-related disulfide bridge with the preexisting Cys 53 on the opposite subunit. We found that the introduction of this disulfide linkage stabilized the mutants A104T and M26I against thermal denaturation, improved their ability to scavenge reactive oxygen species (ROS), and restored a chaperone-like function of blocking alpha-synuclein aggregation. The L166P mutant was far too unstable to be rescued by introduction of the V51C mutation. The results presented here point to the possible development of pharmacological chaperones, which may eventually lead to PD therapeutics.
Logan
Todd
T
Center for Neurologic Diseases, Brigham and Women's Hospital, Boston,Massachusetts 02115, USA.
Clark
Lindsay
L
Ray
Soumya S
SS
D003545:Cysteine / Q000737:chemistry / Q000235:genetics; D030342:Genetic Diseases, Inborn / Q000235:genetics; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D009154:Mutation; D015513:Oncogene Proteins; D010300:Parkinson Disease / Q000235:genetics* / Q000378:metabolism*; D000071617:Protein Deglycase DJ-1; D017434:Protein Structure, Tertiary / Q000235:genetics; D011506:Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D051844:alpha-Synuclein / Q000235:genetics / Q000378:metabolism / Q000502:physiology
D047908:Intracellular Signaling Peptides and Proteins; D018832:Molecular Chaperones; D015513:Oncogene Proteins; D011506:Proteins; D051844:alpha-Synuclein; C105131:PARK7 protein, human; D000071617:Protein Deglycase DJ-1; D003545:Cysteine