Gas chromatography mass spectrometry–based metabolite profiling in plants

Lisec, Jan; Schauer, Nicolas; Kopka, Joachim; Willmitzer, Lothar; Fernie, Alisdair R.

doi:10.1038/nprot.2006.59

Cited by 1,780 publications

(1,446 citation statements)

References 60 publications

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“…(2009) and Lisec et al . (2006). Polar phase extracts were dried using a centrifugal vacuum concentrator and spiked with 100 μ L of an internal standard solution containing 200 μ g mL −1 succinic‐d4 acid, glycine‐d5 and malonic‐d2 acid before analysis.…”

Section: Methodsmentioning

confidence: 99%

Acclimation of metabolism to light in Arabidopsis thaliana: the glucose 6‐phosphate/phosphate translocator GPT2 directs metabolic acclimation

Dyson

Allwood

Feil

et al. 2015

Plant Cell & Environment

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Mature leaves of plants transferred from low to high light typically increase their photosynthetic capacity. In A rabidopsis thaliana, this dynamic acclimation requires expression of GPT2, a glucose 6‐phosphate/phosphate translocator. Here, we examine the impact of GPT2 on leaf metabolism and photosynthesis. Plants of wild type and of a GPT2 knockout (gpt2.2) grown under low light achieved the same photosynthetic rate despite having different metabolic and transcriptomic strategies. Immediately upon transfer to high light, gpt2.2 plants showed a higher rate of photosynthesis than wild‐type plants (35%); however, over subsequent days, wild‐type plants acclimated photosynthetic capacity, increasing the photosynthesis rate by 100% after 7 d. Wild‐type plants accumulated more starch than gpt2.2 plants throughout acclimation. We suggest that GPT2 activity results in the net import of glucose 6‐phosphate from cytosol to chloroplast, increasing starch synthesis. There was clear acclimation of metabolism, with short‐term changes typically being reversed as plants acclimated. Distinct responses to light were observed in wild‐type and gpt2.2 leaves. Significantly higher levels of sugar phosphates were observed in gpt2.2. We suggest that GPT2 alters the distribution of metabolites between compartments and that this plays an essential role in allowing the cell to interpret environmental signals.

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Section: Methodsmentioning

confidence: 99%

Acclimation of metabolism to light in Arabidopsis thaliana: the glucose 6‐phosphate/phosphate translocator GPT2 directs metabolic acclimation

Dyson

Allwood

Feil

et al. 2015

Plant Cell & Environment

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“…The metabolite profiling for primary metabolites was conducted using an established GC-MS protocol as described [30]. Chromatograms and mass spectra were evaluated by Chroma TOF Ò 4.2 (Leco, St. Joseph, MI) and TagFinder 4.0 [31] for quantification and annotation of the peaks using the MPI Golm Metabolome Database (GMD, http://gmd.mpimp-golm.mpg.de/, [32]).…”

Section: Metabolome Analysismentioning

confidence: 99%

Molecular mechanisms of desiccation tolerance in the resurrection glacial relic Haberlea rhodopensis

Gechev¹,

Benina²,

Obata

et al. 2012

Cell. Mol. Life Sci.

165

243

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“…An aliquot of 200 µL was taken and stored at −80°C until GC-and LC-MS analysis. Derivatization and analyses of metabolites by a GC-MS 7890A mass spectrometer (Agilent, Santa Clara, CA) were carried out as described 297 . Metabolites were identified in comparison to Metabolomic Discoveries' database entries of authentic standards.…”

Section: Non-targeted Metabolite Profilingmentioning

confidence: 99%

Tailoring recombinant protein quality by rational media design

Brühlmann

Jordan

Hemberger³

et al. 2015

Biotechnology Progress

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Clinical efficacy and safety of recombinant proteins are closely associated with their structural characteristics. The major quality attributes comprise glycosylation, charge variants (oxidation, deamidation, and C‐ & N‐terminal modifications), aggregates, low‐molecular‐weight species (LMW), and misincorporation of amino acids in the protein backbone. Cell culture media design has a great potential to modulate these quality attributes due to the vital role of medium in mammalian cell culture. The purpose of this review is to provide an overview of the way both classical cell culture medium components and novel supplements affect the quality attributes of recombinant therapeutic proteins expressed in mammalian hosts, allowing rational and high‐throughput optimization of mammalian cell culture media. A selection of specific and/or potent inhibitors and activators of oligosaccharide processing as well as components affecting multiple quality attributes are presented. Extensive research efforts in this field show the feasibility of quality engineering through media design, allowing to significantly modulate the protein function. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:615–629, 2015

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Gas chromatography mass spectrometry–based metabolite profiling in plants

Cited by 1,780 publications

References 60 publications

Acclimation of metabolism to light in Arabidopsis thaliana: the glucose 6‐phosphate/phosphate translocator GPT2 directs metabolic acclimation

Acclimation of metabolism to light in Arabidopsis thaliana: the glucose 6‐phosphate/phosphate translocator GPT2 directs metabolic acclimation

Molecular mechanisms of desiccation tolerance in the resurrection glacial relic Haberlea rhodopensis

Tailoring recombinant protein quality by rational media design

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