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Lipid signaling protocols

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Cover of 'Lipid signaling protocols'

Table of Contents

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    Book Overview
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    Chapter 1 Method for Assaying the Lipid Kinase Phosphatidylinositol-5-phosphate 4-kinase α in Quantitative High-Throughput Screening (qHTS) Bioluminescent Format
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    Chapter 2 Assaying Ceramide Synthase Activity In Vitro and in Living Cells Using Liquid Chromatography-Mass Spectrometry
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    Chapter 3 Lipid Signaling Protocols
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    Chapter 4 Identification of the Interactome of a Palmitoylated Membrane Protein, Phosphatidylinositol 4-Kinase Type II Alpha
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    Chapter 5 Measurement of Long-Chain Fatty Acyl-CoA Synthetase Activity
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    Chapter 6 Qualitative and Quantitative In Vitro Analysis of Phosphatidylinositol Phosphatase Substrate Specificity
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    Chapter 7 Luciferase Reporter Assays to Assess Liver X Receptor Transcriptional Activity
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    Chapter 8 Metabolically Biotinylated Reporters for Electron Microscopic Imaging of Cytoplasmic Membrane Microdomains
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    Chapter 9 Fluorescence Recovery After Photobleaching Analysis of the Diffusional Mobility of Plasma Membrane Proteins: HER3 Mobility in Breast Cancer Cell Membranes.
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    Chapter 10 Isolation and Analysis of Detergent-Resistant Membrane Fractions
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    Chapter 11 Detection of Isolated Mitochondria-Associated ER Membranes Using the Sigma-1 Receptor
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    Chapter 12 Using Surface Plasmon Resonance to Quantitatively Assess Lipid–Protein Interactions
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    Chapter 13 Analyzing Protein–Phosphoinositide Interactions with Liposome Flotation Assays
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    Chapter 14 High-Throughput Fluorometric Assay for Membrane–Protein Interaction
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    Chapter 15 Guidelines for the Use of Protein Domains in Acidic Phospholipid Imaging.
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    Chapter 16 Analysis of Sphingolipid Synthesis and Transport by Metabolic Labeling of Cultured Cells with [ 3 H]Serine
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    Chapter 17 Determination and Characterization of Tetraspanin-Associated Phosphoinositide-4 Kinases in Primary and Neoplastic Liver Cells
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    Chapter 18 Analysis of the Phosphoinositide Composition of Subcellular Membrane Fractions
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    Chapter 19 Single-Molecule Imaging of Signal Transduction via GPI-Anchored Receptors
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    Chapter 20 Measuring Phosphatidylinositol Generation on Biological Membranes
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    Chapter 21 Assay for CDP-Diacylglycerol Generation by CDS in Membrane Fractions
Attention for Chapter 5: Measurement of Long-Chain Fatty Acyl-CoA Synthetase Activity
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Chapter title
Measurement of Long-Chain Fatty Acyl-CoA Synthetase Activity
Chapter number 5
Book title
Lipid Signaling Protocols
Published in
Methods in molecular biology, January 2016
DOI 10.1007/978-1-4939-3170-5_5
Pubmed ID
Book ISBNs
978-1-4939-3169-9, 978-1-4939-3170-5
Authors

Joachim Füllekrug, Margarete Poppelreuther

Abstract

Long-chain fatty acyl-CoA synthetases (ACS) are a family of essential enzymes of lipid metabolism, activating fatty acids by thioesterification with coenzyme A. Fatty acyl-CoA molecules are then readily utilized for the biosynthesis of storage and membrane lipids, or for the generation of energy by ß-oxidation. Acyl-CoAs also function as transcriptional activators, allosteric inhibitors, or precursors for inflammatory mediators. Recent work suggests that ACS enzymes may drive cellular fatty acid uptake by metabolic trapping, and may also regulate the channeling of fatty acids towards specific metabolic pathways. The implication of ACS enzymes in widespread lipid associated diseases like type 2 diabetes has rekindled interest in this protein family. Here, we describe in detail how to measure long-chain fatty acyl-CoA synthetase activity by a straightforward radiometric assay. Cell lysates are incubated with ATP, coenzyme A, Mg(2+), and radiolabeled fatty acid bound to BSA. Differential phase partitioning of fatty acids and acyl-CoAs is exploited to quantify the amount of generated acyl-CoA by scintillation counting. The high sensitivity of this assay also allows the analysis of small samples like patient biopsies.

Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 26 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Spain 1 4%
Unknown 25 96%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 8 31%
Researcher 4 15%
Professor 3 12%
Professor > Associate Professor 2 8%
Student > Master 1 4%
Other 2 8%
Unknown 6 23%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 8 31%
Agricultural and Biological Sciences 6 23%
Immunology and Microbiology 1 4%
Social Sciences 1 4%
Medicine and Dentistry 1 4%
Other 1 4%
Unknown 8 31%