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Full Description
This volume explores methods used to examine the structure-function relationship of glucose transporters both in vitro and ex vivo. Chapters in this book cover topics such as expression and purification of glucose transporters in heterologous expression systems, crystal structure determination, glucose transport activity assays in cellular and synthetic model systems, investigation of the GLUT4 membrane trafficking machinery by novel labeling strategies combined with advanced fluorescence microscopy methods, and tracking GLUT2 translocation by live cell imaging. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and comprehensive, Glucose Transport: Methods and Protocols is a valuable resource that helps researchers with determining specific roles of different GLUTs in various organisms.
Contents
Expression and Purification of Rat Glucose Transporter 1 in Pichia pastoris.- Crystallization and Structural Determination of the Human Glucose Transporters GLUT1 and GLUT3.- Screening and Scale-Up of GLUT Transporter Constructs Suitable for Biochemical and Structural Studies.- GLUT Characterization using Frog Xenopus laevis Oocytes.- Glucose Uptake in Heterologous Expression Systems.- Evaluating the Impact of GLUT Inhibition on Cellular Bioenergetics Using a Seahorse XF Analyzer.- Glucose Transport Activity Measured in Giant Vesicles.- Design, Synthesis, and Evaluation of GLUT Inhibitors.- Applying Microfluidic Systems to Study Effects of Glucose at Single Cell Level.- A Growth-Based Screening System for Hexose Transporters in Yeast.- Identification of Insulin Activated Rab Proteins in Adipose Cells Using Bio-ATB-GTP Photolabeling Technique.- Total Internal Reflection Fluorescence Microscopy to Study GLUT4 Trafficking.- Translocation and Redistribution of GLUT4 Using a Dual-Labeled Reporter Assay.- GLUT4 Translocation in Single Muscle Cells in Culture: Epitope Detection by Immunofluorescence.- Glucose Transport: Methods for Interrogating GLUT4 Trafficking in Adipocytes.- Proximity Ligation Assay to Study the GLUT4 Membrane Trafficking Machinery.- Quantification of Cell Surface Glucose Transporters in the Heart Using a Biotinylated Photolabeled Assay.- Tracking GLUT2 Translocation by Live Cell Imaging.- GLUT2-Expressing Neurons as Glucose Sensors in the Brain: Electrophysiological Analysis.
