Degradation of alpha-synuclein by proteasome.
Mutations in alpha-synuclein are known to be associated with Parkinson's disease (PD). The coexistence of this neuronal protein with ubiquitin and proteasome subunits in Lewy bodies in sporadic disease suggests that alterations of alpha-synuclein catabolism may contribute to the pathogenesis of PD. The degradation pathway of alpha-synuclein has not been identified nor has the kinetics of this process been described. We investigated the degradation kinetics of both wild-type and A53T mutant 6XHis-tagged alpha-synuclein in transiently transfected SH-SY5Y cells. Degradation of both isoforms followed first-order kinetics over 24 h as monitored by the pulse-chase method. However, the t((1)/(2)) of mutant alpha-synuclein was 50% longer than that of the wild-type protein (p < 0.01). The degradation of both recombinant proteins and endogenous alpha-synuclein in these cells was blocked by the selective proteasome inhibitor beta-lactone (40 microM), indicating that both wild-type and A53T mutant alpha-synuclein are degraded by the ubiquitin-proteasome pathway. The slower degradation of mutant alpha-synuclein provides a kinetic basis for its intracellular accumulation, thus favoring its aggregation.
Bennett
M C
MC
Experimental Therapeutics Branch, NINDS, National Institutes of Health, Bethesda, Maryland 20892, USA.
Bishop
J F
JF
Leng
Y
Y
Chock
P B
PB
Chase
T N
TN
Mouradian
M M
MM
D003546:Cysteine Endopeptidases / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D007783:Lactones / Q000494:pharmacology; D009097:Multienzyme Complexes / Q000187:drug effects / Q000378:metabolism*; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex; D011993:Recombinant Fusion Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D051843:Synucleins; D014407:Tumor Cells, Cultured; D051844:alpha-Synuclein
D007783:Lactones; D009097:Multienzyme Complexes; D009419:Nerve Tissue Proteins; D011993:Recombinant Fusion Proteins; C497604:SNCA protein, human; D051843:Synucleins; D051844:alpha-Synuclein; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex