Nutrient Sensing in Eukaryotes

This detailed volume explores various methods through which nutrient sensing can be studied, particularly in multicellular eukaryotes. Beginning with a section on the monitoring of cellular energy status by sensing of adenine nucleotides, the book continues by delving into numerous techniques for researching both intracellular and extracellular nutrient sensing. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Nutrient Sensing in Eukaryotes serves as an ideal guide for researchers aiming to develop related or novel methods to study the sensing of nutrients in their own systems.

Chapter 3 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

Measuring Cellular Adenine Nucleotides by Liquid Chromatography Coupled Mass Spectrometry.- FRET-Based Sensor for Measuring Adenine Nucleotide Binding to AMPK.- Monitoring Cellular Energy Balance in Single Cells Using Fluorescent Biosensors for AMPK.- Assays of Different Mechanisms of AMPK Activation, and Use of Cells Expressing Mutant AMPK to Delineate Activation Mechanisms.- Methods to Determine Lysosomal AMPK Activation.- Sensing of Long Chain Fatty Acyl-CoA Esters by AMPK.- Visualization of Subcellular mTOR Complex 1 Activity with a FRET-Based Sensor (TORCAR).- Determination of Nutrient Ligand - Sensor Binding Affinity.- Amino Acid Sensing by Transceptors: Exploring Substrate-Induced Regulation of Amino Acid Transporters and Transporter Expression.- Purification and Analysis of eIF2a Phosphorylation by Stress-Activated Protein Kinase Gcn2 from S. cerevisiae.- An In Vitro System to Study Mammalian GCN2 Regulation.- Sensing of Cholesterol by Squalene Monooxygenase.- Large Scale Protein Production and Activity Assay Protocols for Human Glycogen Synthase-Glycogenin Complex.- Measurement of Neurotransmitters and Metabolites in Cell Culture and In Vitro Using Genetically Encoded PBP-Based Biosensors.- Calcium Imaging in Hypothalamic Cells.- Nutrient Sensing by Lingual G-Protein Coupled Taste Receptors.