Unknown

Dataset Information

0

Relationships between the sequence of alpha-synuclein and its membrane affinity, fibrillization propensity, and yeast toxicity.


ABSTRACT: To investigate the alpha-synuclein protein and its role in Parkinson's disease, we screened a library of random point mutants both in vitro and in yeast to find variants in an unbiased way that could help us understand the sequence-phenotype relationship. We developed a rapid purification method that allowed us to screen 59 synuclein mutants in vitro and discovered two double-point mutants that fibrillized slowly relative to wild-type, A30P, and A53T alpha-synucleins. The yeast toxicity of all of these proteins was measured, and we found no correlation with fibrillization rate, suggesting that fibrillization is not necessary for synuclein-induced yeast toxicity. We found that beta-synuclein was of intermediate toxicity to yeast, and gamma-synuclein was non-toxic. Co-expression of Parkinson's disease-related genes DJ-1, parkin, Pink1, UCH-L1, or synphilin, with synuclein, did not affect synuclein toxicity. A second screen, of several thousand library clones in yeast, identified 25 non-toxic alpha-synuclein sequence variants. Most of these contained a mutation to either proline or glutamic acid that caused a defect in membrane binding. We hypothesize that yeast toxicity is caused by synuclein binding directly to membranes at levels sufficient to non-specifically disrupt homeostasis.

SUBMITTER: Volles MJ 

PROVIDER: S-EPMC1868670 | biostudies-literature | 2007 Mar

REPOSITORIES: biostudies-literature

altmetric image

Publications

Relationships between the sequence of alpha-synuclein and its membrane affinity, fibrillization propensity, and yeast toxicity.

Volles Michael J MJ   Lansbury Peter T PT  

Journal of molecular biology 20061221 5


To investigate the alpha-synuclein protein and its role in Parkinson's disease, we screened a library of random point mutants both in vitro and in yeast to find variants in an unbiased way that could help us understand the sequence-phenotype relationship. We developed a rapid purification method that allowed us to screen 59 synuclein mutants in vitro and discovered two double-point mutants that fibrillized slowly relative to wild-type, A30P, and A53T alpha-synucleins. The yeast toxicity of all o  ...[more]

Similar Datasets

| S-EPMC7442712 | biostudies-literature
| S-EPMC5485007 | biostudies-literature
| S-EPMC2581432 | biostudies-literature
| S-EPMC7726797 | biostudies-literature
| S-EPMC3551866 | biostudies-literature
2010-01-04 | GSE11633 | GEO
| S-EPMC10357493 | biostudies-literature
| S-EPMC9763379 | biostudies-literature
| S-EPMC2729362 | biostudies-literature
| S-EPMC7075201 | biostudies-literature