PMID: 9425111Feb 28, 1998Paper

Serine-294 and threonine-295 in the exofacial loop domain between helices 7 and 8 of glucose transporters (GLUT) are involved in the conformational alterations during the transport process

The Biochemical Journal
H DoegeH G Joost

Abstract

The role of a conserved polar motif (STS) in the exofacial loop between helices 7 and 8 of GLUT4 for transporter function was investigated by site-directed mutagenesis and expression of the constructs in COS-7 cells. Reconstituted glucose-transport activity, cytochalasin B binding and photolabelling with the exofacial label 2-N4-(1-azi-2,2,2-trifluoroethyl)benzoyl-1, 3-bis-(d-mannosyloxy)-2-propylamine (ATB-BMPA) were assayed in membranes from transfected cells and corrected for immunoreactivity of expressed transporters. Replacement of Ser-294 with Ala or Thr suppressed transport activity and cytochalasin B binding. ATB-BMPA photolabelling was normal in S294A mutants, and even increased in S294T mutants. Replacement of Thr-295 with Ala suppressed transport activity and cytochalasin B binding, whereas ATB-BMPA photolabelling was normal; substitution of Ser failed to alter the investigated parameters. Similarly, exchanging Ser-296 for Ala generated a normally functioning protein. The data suggest that Ser-294 and Thr-295 are involved in the conformational change in GLUT during the transport process, and that their substitution may arrest the transporter in an outward-facing conformation.

Citations

May 24, 2000·The Journal of Biological Chemistry·H DoegeH G Joost
Feb 20, 2014·Proceedings of the National Academy of Sciences of the United States of America·M Gregor MadejH Ronald Kaback
Feb 8, 2000·The Journal of Biological Chemistry·T Kasahara, M Kasahara
Sep 29, 2000·FEBS Letters·J V PlanasJ Gutiérrez
May 4, 2018·Histochemistry and Cell Biology·Sergey Shityakov, Carola Y Förster
Jun 28, 2020·Pflügers Archiv : European journal of physiology·Geoffrey D Holman
Sep 13, 2013·The Biochemical Journal·Ying FengGeoffrey D Holman
Nov 22, 2018·The Biochemical Journal·Geoffrey D Holman
Dec 26, 2008·Journal of Biomolecular Structure & Dynamics·Suma Mohan SGopalakrishnapillai Anilkumar
Jan 24, 2009·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Genciana TerovaMarco Saroglia

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