Analysis of synphilin-1 and synuclein interactions by yeast two-hybrid beta-galactosidase liquid assay.
Synphilin-1 interacts with alpha-synuclein, which has been implicated in the pathogenesis of Parkinson's disease (PD). By examination of their interactions quantitatively, with the use of the yeast two-hybrid beta-galactosidase assay, we find that the synuclein amino acid (aa) 1-65 region is sufficient for an interaction. A central domain of synphilin-1, aa 349-555, is both necessary and sufficient for an interaction with alpha-synuclein. We did not observe an effect of the synuclein A53T mutation, which causes one familial form of PD, on interactions with synphilin-1. However, the A30P mutation caused an increase in the interaction between the synuclein aa 1-65 fragment and the synphilin-1 central domain.
Neystat
Michael
M
Department of Neurology, Columbia University, New York, NY 10032, USA.
Rzhetskaya
Margarita
M
Kholodilov
Nikolai
N
Burke
Robert E
RE
D002352:Carrier Proteins / Q000502:physiology*; D004347:Drug Interactions; D006801:Humans; D009154:Mutation / Q000502:physiology; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000502:physiology*; D020033:Protein Isoforms / Q000502:physiology; D051843:Synucleins; D020798:Two-Hybrid System Techniques; D051844:alpha-Synuclein; D001616:beta-Galactosidase
D002352:Carrier Proteins; D009419:Nerve Tissue Proteins; D020033:Protein Isoforms; C497604:SNCA protein, human; C118958:SNCAIP protein, human; D051843:Synucleins; D051844:alpha-Synuclein; D001616:beta-Galactosidase