Dataset Information

Quantitative proteomics characterization of proteins modified by very long chain fatty acids (VLCFAs) under necroptosis

ABSTRACT: Necroptosis is a caspase-independent form of regulated cell death that is characterized by membrane permeabilization and rupture. This membrane rupture is responsible for the inflammatory properties of necroptosis and is critical for disease states involving this process. In efforts to understand how lipids might contribute to necroptosis, we previously showed that saturated very long chain fatty acids (VLCFAs) are functionally involved in this process, potentially through protein fatty acylation. Here we define the scope of protein acylation by saturated VLCFAs during necroptosis using a clickable lipid analog and IonStar quantitative proteomics.As a result, a total of 1672 proteins were quantified with high precision and no missing data across samples in the same condition. 1267 were detected with higher abundances in control cells and 405 were detected with higher abundance in necroptotic conditions.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell

DISEASE(S): Colon Cancer

SUBMITTER: Shichen Shen

LAB HEAD: Jun Qu

PROVIDER: PXD022527 | Pride | 2021-05-25

REPOSITORIES: Pride

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Publications

Protein acylation by saturated very long chain fatty acids and endocytosis are involved in necroptosis.

Pradhan Apoorva J AJ Lu Daniel D Parisi Laura R LR Shen Shichen S Berhane Ilyas A IA Galster Samuel L SL Bynum Kiana K Monje-Galvan Viviana V Gokcumen Omer O Chemler Sherry R SR Qu Jun J Kay Jason G JG Atilla-Gokcumen G Ekin GE

Cell chemical biology 20210412 9

Necroptosis is a form of cell death characterized by receptor-interacting protein kinase activity and plasma membrane permeabilization via mixed-lineage kinase-like protein (MLKL). This permeabilization is responsible for the inflammatory properties of necroptosis. We previously showed that very long chain fatty acids (VLCFAs) are functionally involved in necroptosis, potentially through protein fatty acylation. Here, we define the scope of protein acylation by saturated VLCFAs during necroptosi ...[more]

PMID: 33848465

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Project description:Neurodegenerative brain disorders become more common in the aged. Most of these disorders are associated with or caused by selective death of certain neuronal subpopulations. The mechanisms underlying the differential vulnerability of certain neuronal populations are still largely unexplored and few neuroprotective treatments are available to date. Elucidation of these mechanisms may lead to a greater understanding of the pathogenesis and treatment of neurodegenerative diseases. Moreover, preconditioning by a short seizure confers neuroprotection following a subsequent prolonged seizure. Our goal is to identify pathways that confer vulnerability and resistance to neurotoxic conditions by comparing the basal and preconditioned gene expression profiles of three differentially vulnerable hippocampal neuron populations. Hippocampal CA1 and CA3 pyramidal neurons are highly susceptible to seizures and ischemia, whereas dentate gyrus granule cells are relatively resistant. A brief preconditioning seizure confers protection to the pyramidal cells. We will first determine gene expression profiles of untreated rat CA1 and CA3 pyramidal cells, and dentate granule cells, using laser capture microscopy to obtain region-specific neuronal mRNA. We will then determine the effect of a brief preconditioning seizure, which is neuroprotective in CA1 and CA3, on these expression profiles. We hypothesize that common molecular mechanisms exist in neurons that determine their susceptibility to seizure-induced injury. Intrinsic differences in gene expression exist between hippocampal glutamatergic CA1 and CA3 pyramidal neurons, on the one hand, and dentate granule cells on the other, which contribute to the greater susceptibility of pyramidal neurons to degeneration in experimental stroke and epilepsy. We specifically hypothesize that differences in basal energy metabolism genes may confer differential susceptibility to neurodegeneration produced by seizures and ischemia. Anesthetized animals will be sacrificed by decapitation, and frozen 10 micron sections will be lightly stained with cresyl violet to identify cell layers in the hippocampus. Approximately 1000 neurons from each of the three cell layers will be isolated by LCM. Poly-A RNA will be amplified using a modified Eberwine protocol. The quality of our aRNA will be evaluated by quantitative RT-PCR of GluR6 and KA2 mRNA levels before we send the samples to the Center for labeling and hybridization to Affymetrix rat 230A arrays. We will provide a one-round amplification cDNA product to the center for labeling and hybridization. This protocol is identical to a previously approved study by Jim Greene in our laboratory.

Dataset Information

Quantitative proteomics characterization of proteins modified by very long chain fatty acids (VLCFAs) under necroptosis

Publications

Protein acylation by saturated very long chain fatty acids and endocytosis are involved in necroptosis.

Similar Datasets

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