Mitoguardin-2–mediated lipid transfer preserves mitochondrial morphology and lipid droplet formation
By savannah onPublished: 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.
Primary data associated with doi: 10.7554/eLife.67900 (Pathogenic LRRK2 control of primary cilia and Hedgehog signaling in neurons and astrocytes of mouse brain)
By quincy.tichenor onImmunofluorescence microscopy images, raw immunoblotting data, and tabular data used to generate graphs shown in all figures.
Mass spectrometry proteomic data supporting the PPM1H/Rab8A crosslinking study described in doi: 10.15252/embr.202152675
By quincy.tichenor onMass spectrometry proteomic data supporting the PPM1H/Rab8A crosslinking study described in doi: 10.15252/embr.202152675, deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD026367
cDNA clone for bacterial expression of human PPM1H H153D
By quincy.tichenor onPlasmid for bacterial expression of human PPM1H (H153D); contains TEV protease site for removal of 6His tag.
Endoplasmic Reticulum Membrane Contact Sites, Lipid Transport, and Neurodegeneration
By quincy.tichenor onReview: Several mutations of genes that encode proteins localized at the endoplasmic reticulum membrane contact sites result in familial neurodegenerative diseases. Here, the authors provide an overview of such diseases, with a specific focus on proteins that directly or indirectly impact lipid transport.
Neuronal hyperactivity–induced oxidant stress promotes in vivo α-synuclein brain spreading
By quincy.tichenor onPublished: This study investigated the relationship between neuronal activity and interneuronal transfer of α-synuclein, a Parkinson-associated protein, and elucidated mechanisms underlying this relationship.
Genetic variations in GBA1 and LRRK2 genes: Biochemical and clinical consequences in Parkinson disease
By quincy.tichenor onReview: This review compares GBA1 and LRRK2-associated PD, and highlights possible genotype-phenotype associations for GBA1 and LRRK2 separately, based on biochemical consequences of single variants.
Code for simulating FSCV artifacts
By quincy.tichenor onCode for simulating FSCV artifacts for algorithm validation in Amjad et al. 2024.
Python script used to generate the heatmap representation of identified LRRK1 phosphosites reported in doi: 10.1042/BCJ20220308
By quincy.tichenor onPython script used to generate the heatmap representation of identified high-confident LRRK1 phosphosites reported in doi: 10.1042/BCJ20220308. It contains the data used for analysis as well as the necessary FASTA sequences for mapping of phosphosite positions on peptides to the original protein sequence.
Primary data associated with Figure 3–Figure Supplement 4 in doi: 10.1101/2022.04.25.489459
By quincy.tichenor onRaw 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").
Primary data associated with doi: 10.1101/2022.04.25.489459 (A Feed-forward Pathway Drives LRRK2 kinase Membrane Recruitment and Apparent Activation)
By quincy.tichenor onRaw immunoblotting data; tabular data for quantifications; immunofluorescence images.
Primary data associated with doi: 10.1042/BCJ20220308 (PKC isoforms activate LRRK1 kinase by phosphorylating conserved residues (Ser1064, Ser1074 and Thr1075) within the CORB GTPase domain)
By quincy.tichenor onRaw immunoblotting data; immunofluorescence microscopy images; tabular data for quantification; autoradiograph films; scans of Coomassie-stained protein gels; Prism files with statistical analysis.
Primary data associated with doi: 10.1042/BCJ20220161 (Impact of 100 LRRK2 variants linked to Parkinson’s Disease on kinase activity and microtubule binding)
By quincy.tichenor onRaw immunoblotting data; tabular data for quantifications; numerical data for in vitro assays; immunofluorescence images; Prism files for statistical analysis.
Preparation and imaging of enriched Golgi from GolgiTAG-IP using Transmission Electron Microscopy
By quincy.tichenor onThis protocol describes how Golgi, isolated from cells by GolgiTAG immunoprecipitation (IP) (as described in dx.doi.org/10.17504/protocols.io.6qpvrdjrogmk/v1), can be prepared and imaged using TEM. This protocol can also be used to image any organelles isolated using various organelle-IP protocols that are available.
Immunofluorescence imaging of cells expressing GolgiTAG (TMEM115-3HA) or HA-tagged non-Golgi annotated proteins
By quincy.tichenor onThis method that can be used to verify the correct localisation of HA-tagged Golgi proteins (like TMEM115-3HA, or GolgiTAG), by assessing their colocalisation with known Golgi markers such as GM130, GOLGIN97 and ACBD3 or can be used to assess if HA-tagged, non-Golgi annotated proteins transiently expressed in cells localise at the Golgi.
Immunoblotting analysis of samples from GolgiTAG (TMEM115-3HA) immunoprecipitation
By quincy.tichenor onThis protocol describes an immunoblotting method to assess efficient enrichment of Golgi proteins in Golgi immunoprecipitation products compared to whole cell lysates and the purity of the immunoprecipitated Golgi by monitoring the expression of other organelles’ markers. This method can also be used to verify the expression of GolgiTAG (TMEM115-3HA) in cells.
Generation of stable cell lines via retroviral or lentiviral transduction
By quincy.tichenor onThis protocol details how to generate stable cell lines using a retrovirus system (can also be used for lentivirus systems) and includes the production of retroviruses encoding the transgene of interest in HEK293FT and the subsequent transduction of the cell line that is intended to stably express the protein of interest.
Crystal structure of Wild-Type PPM1H phosphatase
By quincy.tichenor onCrystal structure of WT PPM1H phosphatase (as reported in 10.15252/embr.202152675) deposited in the Protein Data Bank with code 7L4J.
cDNA clone for bacterial expression of human PPM1H D288A
By quincy.tichenor onPlasmid for bacterial expression of human PPM1H (D288A); contains TEV protease site for removal of 6His tag.
Protein interaction network analysis for Mendelian diseases
By quincy.tichenor onThis protocol describes the steps to use experimentally validated human data to create a protein-protein interaction network (PPIN) based on disease causative genes. Network analysis (combination of topological functional analyses) will lead to the identification of biological processes relevant for disease and disease endophenotypes.