Comparison of quantitatively determined isotopologue patterns of different metabolites resulting from a given experiment allows the reconstruction of labeling patterns of essential intermediates in the major pathways of central metabolism.
In this study, we showed that by monitoring labeling patterns of several key intermediate metabolites in core metabolism, it is possible to quantitate the relative flux ratios for important branch points, such as the malate node.
In the late exponential phase, biomass samples from the cultivations were harvested, hydrolyzed, derivatized and analyzed by gas GC-MS for determination of labelling patterns of intracellular metabolites.
To obtain a quantitative molecular insight into the organization of podosomes and podosome super-structures we have correlated fluorescence labeling patterns of ventral membranes by using light microscopy and high resolution SEM performed on the same samples.
Notably, Western blot labeling patterns of MMP pro- and active subspecies seem not readily translatable between mice and human samples.
Finally, we examined labelling patterns of cPML, an activator of TGFβ signalling that is sequestered in the nucleus, by binding to TGIF.
Notably, quantification of labelling patterns of xylulose-5-P, dihydroxyacetone and glyceraldeyde phosphate, which are directly linked to methanol metabolism, could be measured by first time.
Besides, this approach allowed an improved accuracy for the calculation of fluxes through methanol assimilating pathways, as labelling patterns of some metabolite intermediates of these pathways (DHAP, GA3Pper, PPP intermediates) could be directly accessed.
For example, labeling patterns of aspartate and tricarboxylic acid (TCA) cycle intermediates in human lung cancer cells revealed anaplerosis via activation of pyruvate carboxylase, while in yeast a novel riboneogenesis pathway was confirmed by metabolite labeling.
We hypothesized that if cell wall organization were altered in vti13 root hairs, we would see a change in LM15 labelling when compared with labelling patterns of wild-type root hairs.
For determination of labeling patterns of lactate and amino acids, 50 μl of supernatants were lyophilized, resolved in 50 μl N,N-dimethylformamide (0.1% pyridine) and incubated at 80°C for 30 min.
