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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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Scientific Perspectives: Structural Biology of LRRK2 and its Interaction with Microtubules
Mutations in LRRK2 are linked to Parkinson's disease. LRRK2 regulates membrane trafficking and interacts with microtubules. Recent studies have revealed its cytosolic and microtubule-bound forms using cryo-EM and cryo-ET techniques.
Rab8a expression and purification
Recombinant Rab8a expression and purification protocol as used by the Leschziner and Reck-Peterson Labs
LRRK2 RCKW Protein Purification
Protein purification protocol for tag-less LRRK2RCKW as done by Leschziner and Reck-Peterson Labs. Same protocol can be used to purify LRRK1RCKW as well. Original Protocol by David Snead. Modified by Yu Xuan Lin and Mariusz Matyszewski for…
LRRK2 microtubule sedimentation binding assay
Assay to determine LRRK2 protein binding to microtubules
LRRK2RCKW Widefield fluorescence microtubule binding assay
This assay uses TMR labeled LRRK2 or LRRK1 RCKW to measure binding to microtubules in vitro
LRRK1 expression and purification
Protein purification protocol for full-length LRRK1 as done by Leschziner and Reck-Peterson Labs. Original protocol by Janice M. Reimer and Yu Xuan Lin.
PKC isoforms activate LRRK1 kinase by phosphorylating conserved residues (Ser1064, Ser1074 and Thr1075) within the CORB GTPase domain
Leucine-rich-repeat-kinase 1 (LRRK1) and its homologue LRRK2 are multidomain kinases. Here, the authors study the mechanism controlling LRRK1 activity and reveal a novel unexpected activation mechanism.
Image processing of full-length monomeric LRRK2
This protocol assumes that 2000 to 4000 movies of full-length LRRK2 embedded on vitreous ice are collected with an electron microscope equipped with a direct detector. It focuses primarily on the monomeric population of LRRK2 and has been leading to…
Quick guide to use paceTOMO for cryo-ET data collection from Titan Krios
This quick guide provides key minimal steps for preparing the Titan/SerialEM for the tomogram data collection on lamella or in vitro specimens with a K3 camera. paceTOMO routine is also included for a typical tomogram data collection session. Please…
Preparation of LRRK1 RCKW cryo-EM grids
Protocol used to create LRRK1 RCKW grids for cryo-EM used in Snead, Matyszewski, Dickey et al.
Insect Cell Protocol for LRRK1 and LRRK2 Expression
Protocol for expressing LRRK1 and LRRK2 in insect cells.
A mono- and intralink filter (mi-filter) to reduce false identifications in cross-linking mass spectrometry data
Cross-linking mass spectrometry (XL-MS) has become an indispensable tool for the emerging field of systems structural biology over the recent years. However, the confidence in individual protein–protein interactions (PPIs) depends on the correct…
Preparation of LRRK2 RCKW cryo-EM grids
This is Leschziner's Lab updated protocol for making cryo-EM grids for LRRK2 RCKW. This protocol, when using lower protein concentration, results in better monomer and dimer formation than the old protocol.
Mechanism of human PINK1 activation at the TOM complex in a reconstituted system
The authors demonstrate an essential role of the pore-containing subunit TOM40 and its structurally associated subunits TOM7 and TOM22 for PINK1 activation.