Application of <scp>NMR</scp> Spectroscopy to Assessment of Radiation Dose and Time Lapse

Kang, Chang‐Mo; Hyeon, Jin Seong; Kim, So Ra; Lee, Eun Kyeong; Yun, Hyun Jin; Kim, Sun Young; Chae, Young Kee

doi:10.1002/bkcs.10252

Cited by 2 publications

(4 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We did not observe any other metabolites of the TCA cycle, which might imply the source of succinate was glutamate which had been converted from the input 13 C-glucose. Citrate and malate were the other two intermediates of TCA cycle that have been observed in many of cases [ 25 , 33 , 34 , 36 , 37 , 41 – 43 ], but not in this work.…”

Section: Resultsmentioning

confidence: 53%

See 1 more Smart Citation

Relationship between recombinant protein expression and host metabolome as determined by two-dimensional NMR spectroscopy

2017

Self Cite

View full text Add to dashboard Cite

Escherichia coli has been the most widely used host to produce large amounts of heterologous proteins. However, given an input plasmid DNA, E. coli may produce soluble protein, produce only inclusion bodies, or yield little or no protein at all. Many efforts have been made to surmount these problems, but most of them have involved time-consuming and labor-intensive trial-and-error. We hypothesized that different metabolomic fingerprints might be associated with different protein production outcomes. If so, then it might be possible to change the expression pattern by manipulating the metabolite environment. As a first step in testing this hypothesis, we probed a subset of the intracellular metabolites by partially labeling it with 13C-glucose. We tested 71 genes and identified 17 metabolites by employing the two-dimensional NMR spectroscopy. The statistical analysis showed that there existed the metabolite compositions favoring protein production. We hope that this work would help devise a systematic and predictive approach to the recombinant protein production.

show abstract

Section: Resultsmentioning

confidence: 53%

“…Multivariate analysis was performed with the R statistics software package ( http://www.r-project.org ) or MetaboAnalyst 3.0 [ 35 ]. The script for generating PCA scores plots and biplots by R was first written and provided by Ian Lewis (University of Calgary, Calgary, Canada) and further modified in-house [ 36 , 37 ].…”

Section: Methodsmentioning

confidence: 99%

Relationship between recombinant protein expression and host metabolome as determined by two-dimensional NMR spectroscopy

2017

Self Cite

View full text Add to dashboard Cite

show abstract

“…In the previous study, the amount of unlabeled extracts required for the 2D NMR experiments were marginally met with the cells pooled together from 10 culture plates 23,24 . The current study was focused on finding a suitable 13 C‐labeled input material that could still be enough for the NMR experiments with the cells from one plate.…”

Section: Resultsmentioning

confidence: 99%

“…To take the cell size into account, the total protein can be used as a reference, but we still lacked any evidence that the total metabolite amount is proportional to the total protein. The previous study showed that we achieved better results when we used the number of cells as a normalization reference than the total protein 23,24 . The normalization by the number of cells was intended to equalize the total metabolite amount, so we focused on the relative abundance of each metabolite of each sample.…”

Section: Resultsmentioning

confidence: 99%

Comparison of Three Different Entry Molecules for Probing a Metabolic Response to Radiation Exposure by Two‐Dimensional NMR Spectroscopy

Chae

Kang

2020

Bulletin Korean Chem Soc

Self Cite

View full text Add to dashboard Cite

Humans are continuously exposed to radiation, originating from natural backgrounds or artificial sources. We focused on the X-ray irradiations to which people are exposed during routine medical checkups. We probed the cellular metabolic responses by feeding human umbilical vein endothelial cells with three different 13 C-labeled molecules, which was followed by irradiation with three different doses of X-rays, which was followed by four different incubation periods. We monitored the downstream metabolites via two-dimensional NMR spectroscopy to figure out what input was best suited for our purpose. We found that even though the glucose provided the highest number of metabolized products, the glutamine, which was the lowest, provided as much information as the glucose. Second, the glucose and the glutamine were suitable for the purpose of probing the recovery process, but pyruvate was not suitable. Lastly, 48 h seemed to be needed for the cells to recover from the irradiation shock.

show abstract

Application of NMR Spectroscopy to Assessment of Radiation Dose and Time Lapse

Cited by 2 publications

References 33 publications

Relationship between recombinant protein expression and host metabolome as determined by two-dimensional NMR spectroscopy

Relationship between recombinant protein expression and host metabolome as determined by two-dimensional NMR spectroscopy

Comparison of Three Different Entry Molecules for Probing a Metabolic Response to Radiation Exposure by Two‐Dimensional NMR Spectroscopy

Contact Info

Product

Resources

About