Pharmaceutical Research

, 28:2745

Quantitative Glucose and ATP Sensing in Mammalian Cells

  • Dania C. Liemburg-Apers
  • Hiromi Imamura
  • Marleen Forkink
  • Marco Nooteboom
  • Herman G. Swarts
  • Roland Brock
  • Jan A. M. Smeitink
  • Peter H. G. M. Willems
  • Werner J. H. Koopman
Expert Review

DOI: 10.1007/s11095-011-0492-8

Cite this article as:
Liemburg-Apers, D.C., Imamura, H., Forkink, M. et al. Pharm Res (2011) 28: 2745. doi:10.1007/s11095-011-0492-8

ABSTRACT

The functioning and survival of mammalian cells requires an active energy metabolism. Metabolic dysfunction plays an important role in many human diseases, including diabetes, cancer, inherited mitochondrial disorders, and metabolic syndrome. The monosaccharide glucose constitutes a key source of cellular energy. Following its import across the plasma membrane, glucose is converted into pyruvate by the glycolysis pathway. Pyruvate oxidation supplies substrates for the ATP-generating mitochondrial oxidative phosphorylation (OXPHOS) system. To gain cell-biochemical knowledge about the operation and regulation of the cellular energy metabolism in the healthy and diseased state, quantitative knowledge is required about (changes in) metabolite concentrations under (non) steady-state conditions. This information can, for instance, be used to construct more realistic in silico models of cell metabolism, which facilitates understanding the consequences of metabolic dysfunction as well as on- and off-target effects of mitochondrial drugs. Here we review the current state-of-the-art live-cell quantification of two key cellular metabolites, glucose and ATP, using protein-based sensors. The latter apply the principle of FRET (fluorescence resonance energy transfer) and allow measurements in different cell compartments by fluorescence microscopy. We further summarize the properties and applications of the FRET-based sensors, their calibration, pitfalls, and future perspectives.

KEY WORDS

ATeamfibroblastGLUTsystems biology

ABBREVIATIONS

2-DG

2-Deoxy-D-glucose

A

fluorescence acceptor molecule

AcCoA

acetyl-coenzyme A

ADP

adenoside diphosphate

AMP

adenosine monophosphate

AMPK

AMP-activated protein kinase

ANT

adenine nucleotide translocator

ATP

adenoside triphosphate

D

fluorescence donor molecule

DNP

2,4-Dinitrophenol

ECFP

enhanced cyan fluorescent protein

ER

endoplasmic reticulum

EYFP

enhanced yellow fluorescent protein

FCCP

carbonyl cyanide-p-trifluoromethoxyphenylhydrazone

FRET

fluorescence resonance energy transfer

FS

fractional saturation

G6P

glucose-6-phosphate

GFP

green fluorescent protein

GGBP

glucose galactose-binding protein

GK

glucokinase

GLUT

glucose transporter

HK

hexokinase

IAA

iodoacetate

LDH

lactate dehydrogenase

OFP

orange fluorescent protein

OXPHOS

oxidative phosphorylation

PBP

periplasmic binding protein

PDH

pyruvate dehydrogenase

PFK

phosphofructokinase

PK

pyruvate kinase

PKC

protein kinase C

PM

plasma membrane

PPP

pentose phosphate pathway

ROS

reactive oxygen species

SGLT

sodium-dependent glucose cotransporters

SLO

streptolysin O

SNR

signal-to-noise ratio

TCA

tricarboxylic acid

TPA

phorbol 12-myristate 13-acetate

VDAC

voltage-dependent anion channel

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Dania C. Liemburg-Apers
    • 1
    • 2
    • 3
  • Hiromi Imamura
    • 4
  • Marleen Forkink
    • 1
  • Marco Nooteboom
    • 1
    • 2
    • 3
  • Herman G. Swarts
    • 1
  • Roland Brock
    • 1
  • Jan A. M. Smeitink
    • 2
    • 3
  • Peter H. G. M. Willems
    • 1
    • 3
  • Werner J. H. Koopman
    • 1
    • 3
  1. 1.Department of Biochemistry (286) Nijmegen Centre for Molecular Life SciencesRadboud University Nijmegen Medical CentreNijmegenThe Netherlands
  2. 2.Department of Pediatrics Nijmegen Centre for Mitochondrial DisordersRadboud University Nijmegen Medical CentreNijmegenThe Netherlands
  3. 3.Centre for Systems Biology and BioenergeticsRadboud University Nijmegen Medical CentreNijmegenThe Netherlands
  4. 4.Kyoto UniversityKyotoJapan