Affiliation:
1. Department of Chemistry and Biology Toronto Metropolitan University Toronto Canada
2. Faculty of Medicine Fukuoka University Fukuoka Japan
3. Program in Cell Biology The Hospital for Sick Children Toronto Ontario Canada
Abstract
AbstractElevated blood glucose following a meal is cleared by insulin‐stimulated glucose entry into muscle and fat cells. The hormone increases the amount of the glucose transporter GLUT4 at the plasma membrane in these tissues at the expense of preformed intracellular pools. In addition, muscle contraction also increases glucose uptake via a gain in GLUT4 at the plasma membrane. Regulation of GLUT4 levels at the cell surface could arise from alterations in the rate of its exocytosis, endocytosis, or both. Hence, methods that can independently measure these traffic parameters for GLUT4 are essential to understanding the mechanism of regulation of membrane traffic of the transporter. Here, we describe cell population–based assays to measure the steady‐state levels of GLUT4 at the cell surface, as well as to separately measure the rates of GLUT4 endocytosis and endocytosis. © 2023 Wiley Periodicals LLC.Basic Protocol 1: Measuring steady‐state cell surface GLUT4mycBasic Protocol 2: Measuring steady‐state cell surface GLUT4‐HABasic Protocol 3: Measuring GLUT4myc endocytosisBasic Protocol 4: Measuring GLUT4myc exocytosis
Subject
Medical Laboratory Technology,Health Informatics,General Pharmacology, Toxicology and Pharmaceutics,General Immunology and Microbiology,General Biochemistry, Genetics and Molecular Biology,General Neuroscience