Glucose and glycerol transport in adipocytes from a structural perspective

Peng Huang

Research output: ThesisDoctoral Thesis (compilation)

72 Downloads (Pure)

Abstract

Adipocytes are crucial energy reservoirs to maintain metabolic homeostasis of glucose and lipids in the human body.
Glucose transporters (GLUTs) and aquaporins (AQPs) play an important role in metabolic regulation of glucose and
lipids in human adipocytes. Specifically, glucose transporter 4 (GLUT4) and aquaporin 7 (AQP7) are the central
players for glucose transport and glycerol efflux in adipocytes. In addition, GLUT family members are overexpressed
in a vast majority of cancer cells to satisfy their increased energy demand, thus, inhibitors targeting GLUTs are
becoming relevant therapeutics for cancers treatment. To control the uptake/release of nutrients, GLUTs and AQPs
have been suggested to be controlled by trafficking mechanisms. TUG (tether containing UBX domain for GLUT4
in mouse) and PLIN1 (human perilipin 1) have previously been suggested to bind with GLUT4 and AQP7
intracellularly and release them upon hormonal stimulation. Here, the mRNA expression levels of GLUTs and AQPs
in adipose tissue has been investigated, and detailed characterization of the interaction between GLUT4 and ASPL
(human TUG homolog) and AQP7 and PLIN1 in vitro have been executed. In addition, a new series of glucose
transporter 1 (GLUT1) inhibitors was structurally and functionally investigated. Finally, the AQP7 structure was
elucidated by single particle cryo-EM.
In this thesis I suggest that GLUT4 interacts with ASPL using its intracellular helical domain to bind to the Cterminus
of ASPL. Rat GLUT4 was expressed in Pichia pastoris, and purified, and showing even single-particle
distribution in negative staining, providing insight on further structural study of GLUT4 by single particle cryo-EM.
Docking models of the complex of GLUT4 and ASPL were generated and suggest that ASPL forms complex with
GLUT4 by multiple domains including both ubiquitin-like domain (UBL2) and ubiquitin domain (UBX). In addition,
PGL13 and PGL14, as new series of GLUT1 inhibitors were suggested to utilize two sites of GLUT1, the
transmembrane domain and intracellular helical domain. Moreover, in human adipocytes AQP7 gene showed
markedly higher-level expression than other aquaglyceroporins. The C-terminal domain of PLIN1 was suggested
to be central for the complex formation with AQP7 and AQP3. The AQP7 structure was determined at the
resolution of 2.55 Å by cryo-EM, adopting the formation of dimer of tetramers. Two tetramers were dimerized by
extracellular protruding C loops with a rotation of approximate 11° around central axis. The central pore is formed
by four monomers and restricted by two leucine filters. Moreover, well-defined densities were discovered in the
central pores showing decent fitting with some small metabolic products.
Original languageEnglish
QualificationDoctor
Awarding Institution
  • Department of Experimental Medical Science
Supervisors/Advisors
  • Lindkvist, Karin, Supervisor
  • Gourdon, Pontus, Assistant supervisor
Award date2022 Jan 27
Place of PublicationLund
Publisher
ISBN (Print)978-91-8021-169-7
Publication statusPublished - 2022

Bibliographical note

Defense details
Date: 2022-01-27
Time: 13:00
Place: Segerfalksalen, BMC A10, Sölvegatan 17 i Lund. Join by Zoom: https://lu-se.zoom.us/j/69869391433
External reviewer (s)
Name: Drew, David
Title: Professor
Affiliation: Stockholm University

Subject classification (UKÄ)

  • Cell and Molecular Biology

Free keywords

  • adipocytes
  • glucose and glycerol metabolism
  • glucose transporters
  • aquaglyceroporins
  • glucose transporter 4
  • aquaglyceroporin 7
  • protein and protein interaction
  • structure
  • single particle cryo-EM

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