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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.
Article
ER-lysosome lipid transfer protein VPS13C/PARK23 prevents aberrant mtDNA-dependent STING signaling
Published: Mutations in VPS13C cause early onset, autosomal recessive PD. VPS13C encodes a lipid transfer protein that is localized to ER-endosome/lysosome contact sites. The authors demonstrate that depletion of VPS13C causes an accumulation of lysosomes and activated STING, suggesting a link between ER-lysosome lipid transfer and innate immune activation. View original preprint.
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Protocol
Cell culture on EM grids and fluorescence microscopy imaging
This protocol describes the general procedure of seeding mammalian cells on EM grids and confocal fluorescence microscopy imaging of grids for subsequent cryo-tomography experiments, performed.
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Regular maintenance of human pluripotent stem cells
This protocol describes the regular maintenance and passaging human iPSCs.
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Article
Mitoguardin-2–mediated lipid transfer preserves mitochondrial morphology and lipid droplet formation
Published: Lipid transport at membrane contact sites is a critical biological process. The authors identify mitoguardin-2, a mitochondrial protein that functions as a lipid transporter at contact sites between the ER and/or lipid droplets. They show that mitoguardin-2 transfers glycerophospholipids between membranes in vitro.
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Lab Resource
mCh-Miro1(∆TM)-SspB
mCh-Miro1(∆TM)-SspB plasmid has a deletion of transmembrane domain and is used for improved light inducible dimer assay.
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Protocol
RNA isolation and qRT-PCR
This protocol describes the RNA isolation from cultured cells and quantitative RT PCR.
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Transfection and protein over-expression
This protocol provides the details for how to transfect and harvest 3X-FLAG-SHIP164Δ901-1099.
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VPS13C^halo∆1235-1748
Express VPS13C truncation mutant in mammalian cells (VPS13C^halo∆1235-1748).
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Surface protein biotinylation
This protocol describes surface protein labeling with biotin using EZ-Link Sulfo-NHSLC-Biotin. This chemical reacts with primary amines such as lysine but does not permeate cell membranes because of the charge. Thus, it only biotinylates surface proteins.
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Subtomogram averaging and classification
This protocol describes the procedure of subtomogram averaging and 3D classification of VPS13C rod-shaped densities inside cryotomograms of mammalian cells. Sub-tomogram averaging package i3 was used for 3D alignment and classification.
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Article
In situ architecture of the lipid transport protein VPS13C at ER-lysosomes membrane contacts
Publication: Loss-of-function mutations in VPS13C are responsible for rare cases of familial early onset Parkinson’s disease. Using cryo-ET, the authors provide in-situ evidence for a bridge-model of VPS13 in lipid transport. View the original preprint.
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Crystallization, data collection and structure solution of PxP-VAB
Protocol lists the steps to obtain PxP(Mcp1ct)-VABct crystals, X-ray diffraction data acquisition and obtaining the structure solution using molecular replacement with a model generated by AlphaFold2.
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