Can stable isotope mass spectrometry replace ‎radiolabelled approaches in metabolic studies?
Metabolic pathways and the key regulatory points thereof can be deduced using isotopically labelled substrates. One prerequisite is the accurate measurement of the labeling pattern of targeted metabolites. The subsequent estimation of metabolic fluxes following incubation in radiolabelled substrates has been extensively used. Radiolabelling is a sensitive approach and allows determination of total label uptake since the total radiolabel content is easy to detect. However, the incubation of cells, tissues or the whole plant in a stable isotope enriched environment and the use of either mass spectrometry or nuclear magnetic resonance techniques to determine label incorporation within specific metabolites offers the possibility to readily obtain metabolic information with higher resolution. It additionally also offers an important complement to other post-genomic strategies such as metabolite profiling providing insights into the regulation of the metabolic network and thus allowing a more thorough description of plant cellular function. Thus, although safety concerns mean that stable isotope feeding is generally preferred, the techniques are in truth highly complementary and application of both approaches in tandem currently probably provides the best route towards a comprehensive understanding of plant cellular metabolism.
Max Planck Partner Group at the Departamento de Biologia Vegetal, Universidade Federal de Viçosa, 36570-900, Viçosa-MG, Brazil. Electronic address: williambatistadasilva@gmail.com.
Willian
W
Batista Silva
Max-Planck-Institute of Molecular Plant Physiology Am Mühlenberg 1, 14476,Golm Potsdam, Germany. Electronic address: daloso@mpimp-golm.mpg.de.
Danilo M
DM
Daloso
Max-Planck-Institute of Molecular Plant Physiology Am Mühlenberg 1, 14476,Golm Potsdam, Germany. Electronic address: fernie@mpimp-golm.mpg.de.
Alisdair R
AR
Fernie
Max Planck Partner Group at the Departamento de Biologia Vegetal, Universidade Federal de Viçosa, 36570-900, Viçosa-MG, Brazil. Electronic address: nunesnesi@ufv.br.
Adriano
A
Nunes-Nesi
Max Planck Partner Group at the Departamento de Biologia Vegetal, Universidade Federal de Viçosa, 36570-900, Viçosa-MG, Brazil. Electronic address: wlaraujo@ufv.br.
Wagner L
WL
Araújo
D013058:Mass Spectrometry / Q000379:methods; D053858:Metabolic Networks and Pathways; D055432:Metabolomics / Q000379:methods* / Q000639:trends; D010944:Plants / Q000378:metabolism*; D011868:Radioisotopes; D013194:Staining and Labeling
D011868:Radioisotopes
Flux profiling; Isotopic labelling; Metabolic flux analysis; Plant systems biology; Primary metabolism