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research article

Unidirectional steady state rates of central metabolism enzymes measured simultaneously in a living plant tissue

Roscher, A
•
Emsley, L  
•
Raymond, P
Show more
1998
Journal of Biological Chemistry

The unidirectional steady state reaction rates of several enzymes and metabolic fluxes of distinct processes were measured simultaneously in hypoxic maize root tips using two-dimensional phosphorus NMR exchange spectroscopy. A single spectrum monitors ATP synthesis and hydrolysis as well as the activities of four enzymes involved in key pathways of central metabolism: UDP-glucose pyrophosphorylase, phosphoglucomutase, hexose-phosphate isomerase, and enolase, The corresponding unidirectional reaction rates and net metabolic fluxes were calculated from spectral intensities. This method provides a unique picture, at enzyme resolution, of how metabolism reacts in a concerted fashion to changes in external parameters such as temperature and oxygen concentration, By increasing hypoxia via an increase in temperature, we measured the expected increase in glycolysis through enolase activity while total ATP synthesis settled. At the same time, we observed a net flux through phosphoglucomutase and UDP-glucose pyrophosphorylase toward carbohydrate synthesis. This result is discussed in relation to the current hypothesis on the turnover of cell walls and sucrose. This reaction also produces a net flux of pyrophosphate, which is needed by pyrophosphate:fructose-6-phosphate 1-phosphotransferase to work as a glycolytic enzyme.

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Type
research article
DOI
10.1074/jbc.273.39.25053
Web of Science ID

WOS:000076085400012

Author(s)
Roscher, A
Emsley, L  
Raymond, P
Roby, C
Date Issued

1998

Publisher

AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC

Published in
Journal of Biological Chemistry
Volume

273

Issue

39

Start page

25053

End page

25061

Editorial or Peer reviewed

REVIEWED

Written at

OTHER

EPFL units
LRM  
Available on Infoscience
January 8, 2015
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/110152
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