Transfected synphilin-1 forms cytoplasmic inclusions in HEK293 cells.
The discovery of mutations in the gene for alpha-synuclein in familial Parkinson's disease (PD) has led to an increased interest in this pre-synaptic protein. Synphilin-1, a potential synuclein-binding protein, was cloned using yeast two-hybrid assays. The function of synphilin-1 is currently unknown, although it has been reported to be present along with alpha-synuclein in Lewy bodies in PD. In the present study, we monitored synphilin-1 aggregation directly using fusion proteins of synphilin-1 and green fluorescent protein (EGFP). Transfection of synphilin-EGFP fusion proteins formed cytoplasmic inclusions in HEK293 cells. Although these inclusions overlapped with the distribution of alpha-synuclein, they were unlike Lewy bodies in that they were not eosinophilic, and instead were membrane-bound, lipid-rich cytoplasmic inclusions.
O'Farrell
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C
Neurogenetics Laboratory, Mayo Clinic Jacksonville, Jacksonville, FL 32224, USA.
Murphy
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DD
Petrucelli
L
L
Singleton
A B
AB
Hussey
J
J
Farrer
M
M
Hardy
J
J
Dickson
D W
DW
Cookson
M R
MR
D000595:Amino Acid Sequence; D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics; D002460:Cell Line / Q000737:chemistry / Q000648:ultrastructure; D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases / Q000378:metabolism; D004622:Embryo, Mammalian; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D006801:Humans; D002479:Inclusion Bodies / Q000737:chemistry*; D007668:Kidney; D007976:Leupeptins / Q000494:pharmacology; D016631:Lewy Bodies / Q000737:chemistry / Q000648:ultrastructure; D008164:Luminescent Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000235:genetics; D010300:Parkinson Disease / Q000235:genetics / Q000473:pathology; D017354:Point Mutation; D011480:Protease Inhibitors / Q000494:pharmacology; D046988:Proteasome Endopeptidase Complex; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D014162:Transfection; D020798:Two-Hybrid System Techniques
D002352:Carrier Proteins; D007976:Leupeptins; D008164:Luminescent Proteins; D009097:Multienzyme Complexes; D009419:Nerve Tissue Proteins; D011480:Protease Inhibitors; D011993:Recombinant Fusion Proteins; C118958:SNCAIP protein, human; D049452:Green Fluorescent Proteins; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde