A5004693112- Cellular transport and secretion
- Retinal Development and Disorders
- Heat shock proteins research
- thermodynamics and calorimetric analyses
- Photosynthetic Processes and Mechanisms
- Cell Image Analysis Techniques
- Microtubule and mitosis dynamics
- Parkinson's Disease Mechanisms and Treatments
- Plant Reproductive Biology
- Cell death mechanisms and regulation
- Ubiquitin and proteasome pathways
- Congenital heart defects research
- Microbial Inactivation Methods
- Genetic and Kidney Cyst Diseases
- Genetics and Neurodevelopmental Disorders
- Lysosomal Storage Disorders Research
- Advanced Fluorescence Microscopy Techniques
- Protein Kinase Regulation and GTPase Signaling
- Bacterial Genetics and Biotechnology
- Calcium signaling and nucleotide metabolism
- Toxin Mechanisms and Immunotoxins
Stanford University
2020-2022
Research Network (United States)
2022
University of California, Los Angeles
2015
Activating mutations in the leucine-rich repeat kinase 2 (LRRK2) cause Parkinson's disease, and previously we showed that activated LRRK2 phosphorylates a subset of Rab GTPases (Steger et al., 2017). Moreover, Golgi-associated Rab29 can recruit to surface Golgi activate it there for both auto- substrate phosphorylation. Here, define precise binding region Armadillo domain between residues 360-450 show this domain, termed 'site #1,' also bind additional substrates, Rab8A Rab10. identify...
Mutations that enhance LRRK2 protein kinase activity cause inherited Parkinson's disease. phosphorylates a group of Rab GTPase proteins, including Rab10 and Rab12, within the effector-binding switch-II motif. Previous work has indicated PARK16 locus, which harbors gene encoding for Rab29, is involved in Parkinson's, Rab29 operates common pathway with LRRK2. Co-expression stimulates by recruiting to surface trans Golgi network. Here, we report knock-out does not influence endogenous activity,...
Activating mutations in LRRK2 kinase causes Parkinson’s disease. Pathogenic phosphorylates a subset of Rab GTPases and blocks ciliogenesis. Thus, defining novel phospho-Rab interacting partners is critical to our understanding the molecular basis pathogenesis. RILPL2 binds with strong preference LRRK2-phosphorylated Rab8A Rab10. binding partner motor protein effector, Myosin Va. We show here that globular tail domain Va also contains high affinity site for In presence pathogenic LRRK2, MyoVa...
Mid1 and Mid2 are ubiquitin ligases that regulate microtubule dynamics whose mutation is associated with X-linked developmental disorders. We show astrin, a microtubule-organizing protein, co-purifies Mid2, has an overlapping localization at intercellular bridge microtubules, ubiquitinated by on lysine 409, degraded during cytokinesis. depletion led to astrin stabilization cytokinesis, cytokinetic defects, multinucleated cells, cell death. Similarly, expression of K409A mutant in...
Microscale thermophoresis (MST) is a powerful tool to measure the affinities of interactions between proteins. We present here our method for determining binding Rab29 GTPase LRRK2 (Leucine rich repeat kinase 2) N-terminal Armadillo domain. Work from several labs has shown that can recruit Golgi, where it normally resides, or other compartments, when artificially relocalized another cellular compartment. MST enabled us define precise site Rab GTPases on
Abstract Activating mutations in the Leucine Rich Repeat Kinase 2 (LRRK2) cause Parkinson’s disease and previously we showed that activated LRRK2 phosphorylates a subset of Rab GTPases (Steger et al., 2017). Moreover, Golgi-associated Rab29 can recruit to surface Golgi activate it there for both auto- substrate phosphorylation. Here define precise binding region Armadillo domain between residues 360-450 show this domain, termed “Site #1”, also bind additional substrates, Rab8A Rab10....
Abstract Activating mutations in LRRK2 kinase cause Parkinson’s disease. Pathogenic phosphorylates a subset of Rab GTPases and blocks ciliogenesis. Thus, defining novel phospho-Rab interacting partners is critical to our understanding the molecular basis pathogenesis. RILPL2 binds with strong preference LRRK2-phosphorylated Rab8A Rab10. binding partner motor protein effector, Myosin Va. We show here that globular tail domain Va also contains high affinity site for Rab10, certain...
Abstract Mutations that enhance LRRK2 protein kinase activity cause inherited Parkinson’s disease. phosphorylates a group of Rab GTPase proteins, including Rab10 and Rab12, within the effector-binding switch-II motif. Previous work has indicated PARK16 locus, which harbors gene encoding for Rab29, is involved in Parkinson’s, Rab29 operates common pathway with LRRK2. Co-expression stimulates by recruiting to surface trans Golgi network. Here we report knock-out does not influence endogenous...
Several labs have shown that Rab29 GTPase can recruit LRRK2 to the surface of Golgi complex. We describe here a method monitor direct binding N-terminal Armadillo domain using Microscale Thermophoresis. This utilizes purified, His-tagged residues 1-552-labeled with NanoTemper Second Gen NHS-Red 647 dye and bacterially expressed protein. Binding is detected Nanotemper Monolith NT-115 instrument.
Supported lipid bilayers have emerged as an ideal model system to study the interaction of proteins with cellular membranes. We describe here a method monitor recruitment purified LRRK2 kinase onto planar containing lipid-anchored Rab10 protein using Total Internal Reflection Fluorescence (TIRF) Microscopy.This utilizes purified, FLAG-tagged, full length labeled CF633 succinimidyl ester (Biotium) and bacterially expressed eGFP-Rab10-His tagged protein. is captured in real time at 25°C Nikon...
Microscale thermophoresis (MST) is a powerful tool to measure the affinities of interactions between proteins. We present here our method for determining binding Rab29 GTPase LRRK2 (Leucine rich repeat kinase 2) N-terminal Armadillo domain. Work from several labs has shown that can recruit Golgi, where it normally resides, or other compartments, when artificially relocalized another cellular compartment. MST enabled us define precise site Rab GTPases on
Microscale thermophoresis (MST) is a powerful tool to measure the affinities of interactions between proteins. We present here our method for determining binding Rab29 GTPase LRRK2 (Leucine rich repeat kinase 2) N-terminal Armadillo domain. Work from several labs has shown that can recruit Golgi, where it normally resides, or other compartments, when artificially relocalized another cellular compartment. MST enabled us define precise site Rab GTPases on