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Output Catalog
ASAP is committed to accelerating the pace of discovery and informing a path to a cure for Parkinson’s disease through collaboration, research-enabling resources, and data sharing. We’ve created this catalog to showcase the research outputs and tools developed by ASAP-funded programs.
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End-product inhibition of the LRRK2-counteracting PPM1H phosphatase
PPM1H phosphatase reverses Rab GTPase phosphorylation by LRRK2 in Parkinson's disease. PPM1H binds Rab8A and Rab10 at an allosteric site, inhibiting phosphatase activity. Targeting this site may offer a therapeutic strategy.
The bridge-like lipid transport protein VPS13C/PARK23 mediates ER-lysosome contacts upon lysosome damage
Genetic studies link lysosome dysfunction to Parkinson's disease. VPS13C, a PD gene, relocates to damaged lysosomes, aiding in membrane repair. This process involves Rab7 and suggests early lipid transport as a protective response.
Tissue Mito-IP: Immunopurification of Mitochondria from MitoTag Mice
A novel method called 'Mito-IP' enables rapid immunopurification of pure mitochondria in 10 minutes using a chimeric protein with HA tags, improving organelle isolation for various analyses.
PPM1M, a LRRK2-counteracting, phosphoRab12-preferring phosphatase with potential link to Parkinson’s disease
LRRK2 phosphorylates Rab GTPases linked to Parkinson's disease. PPM1M identified as a phosphatase reversing Rab phosphorylation, crucial in LRRK2 pathway. PPM1M mutation found in Parkinson's patients, suggesting a new therapeutic target.
Phosphatome-wide siRNA screen in 3T3 cells
Protocol details a phosphatome-wide siRNA screen in 3T3 cells to identify phosphatases counteracting LRRK2 for pRab12 and pRab10.
Lysosomal GCase (glucocerebrosidase) activity assay
Method to measure lysosomal GBA1 enzyme activity using 4-MUG substrate at low pH to minimize non-lysosomal GCase interference. GBA1 activity higher in purified lysosomes than whole cell extract, confirmed by CBE inhibitor abolishing 4-MUG hydrolysis.
Data upload for Tagless LysoIP method for molecular profiling of lysosomal content in clinical samples
Summary: Raw data for the tagless LysoIP manuscript is available.
PPM1M knockout Mouse Embryonic Fibroblasts (mouse)
PPM1M knockout mouse embryonic fibroblasts compared to wild-type littermate controls.
Lysosomal Glucocerebrosidase is needed for ciliary Hedgehog signaling: A convergent pathway to Parkinson’s disease
Mutations in LRRK2 and GBA1 are common in familial Parkinson’s disease. These mutations inhibit primary cilia formation, blocking Hedgehog signaling and reducing neuroprotective factors, contributing to PD pathogenesis.
Primary data associated with Figure 3–Figure Supplement 4 in doi: 10.1101/2022.04.25.489459
Raw immunoblotting data and tabular data for quantitation used to generate Figure 3–Figure Supplement 4 in doi: 10.1101/2022.04.25.489459 ("A Feed-forward Pathway Drives LRRK2 kinase Membrane Recruitment and Activation").
Binding of Rab29 to the LRRK2 Armadillo domain by Microscale Thermophoresis
Microscale thermophoresis is a powerful tool to measure the affinities of interactions between proteins. We present here our method for determining the binding of Rab29 GTPase to the LRRK2 N-terminal Armadillo domain.
Primary data associated with doi: 10.1042/BCJ20220161 (Impact of 100 LRRK2 variants linked to Parkinson’s Disease on kinase activity and microtubule binding)
Raw immunoblotting data; tabular data for quantifications; numerical data for in vitro assays; immunofluorescence images; Prism files for statistical analysis.
HEK293 TMEM115-3xHA GolgiTAG (CVCL_C8A6)
Cell line generated from HEK293 cells (Cat# CRL-1573, RRID:CVCL_0045) virally transfected with a construct expressing TMEM115-3xHA (GolgiTAG)
Role of autophagy pathway in Parkinson’s disease and related Genetic Neurological disorders
The authors provide a comprehensive overview of the general importance of autophagy in Parkinson’s disease (PD) and related disorders of the central nervous system (CNS).
Staining of cells with GolgiTracker for Golgi flow cytometry analysis
Method that allows the staining of intact Golgi using GolgiTracker for subsequent flow cytometry analysis.