Characterisation of G418-induced metabolic load in recombinant CHO and BHK cells: effect on the activity and expression of central metabolic enzymes

Research output: Contribution to journalJournal articlepeer-review

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Characterisation of G418-induced metabolic load in recombinant CHO and BHK cells : effect on the activity and expression of central metabolic enzymes. / Yallop, C. A.; Nørby, P. L.; Jensen, R.; Reinbach, H.; Svendsen, I.

In: Cytotechnology, Vol. 42, No. 2, 2003, p. 87-99.

Research output: Contribution to journalJournal articlepeer-review

Harvard

Yallop, CA, Nørby, PL, Jensen, R, Reinbach, H & Svendsen, I 2003, 'Characterisation of G418-induced metabolic load in recombinant CHO and BHK cells: effect on the activity and expression of central metabolic enzymes', Cytotechnology, vol. 42, no. 2, pp. 87-99. https://doi.org/10.1023/B:CYTO.0000009821.82741.8c

APA

Yallop, C. A., Nørby, P. L., Jensen, R., Reinbach, H., & Svendsen, I. (2003). Characterisation of G418-induced metabolic load in recombinant CHO and BHK cells: effect on the activity and expression of central metabolic enzymes. Cytotechnology, 42(2), 87-99. https://doi.org/10.1023/B:CYTO.0000009821.82741.8c

Vancouver

Yallop CA, Nørby PL, Jensen R, Reinbach H, Svendsen I. Characterisation of G418-induced metabolic load in recombinant CHO and BHK cells: effect on the activity and expression of central metabolic enzymes. Cytotechnology. 2003;42(2):87-99. https://doi.org/10.1023/B:CYTO.0000009821.82741.8c

Author

Yallop, C. A. ; Nørby, P. L. ; Jensen, R. ; Reinbach, H. ; Svendsen, I. / Characterisation of G418-induced metabolic load in recombinant CHO and BHK cells : effect on the activity and expression of central metabolic enzymes. In: Cytotechnology. 2003 ; Vol. 42, No. 2. pp. 87-99.

Bibtex

@article{d84e62a2be70479cb99b1599e724e282,
title = "Characterisation of G418-induced metabolic load in recombinant CHO and BHK cells: effect on the activity and expression of central metabolic enzymes",
abstract = "In a previous article (Yallop and Svendsen 2001), recombinant CHO and BHK cell lines, expressing the human glucagon receptor and the gastric inhibitory peptide receptor, respectively, showed reduced growth rates and altered nutrient utilisation when grown with increasing concentrations of G418. This response was associated with an increased expression of the neor protein, while expression of the recombinant membrane receptors remained unaltered. The metabolic response was characterised in both cell lines by an increase in the specific rate of glutamine utilisation and in CHO cells by a decrease in the yield of lactate from glucose, suggesting a change in the flux of glucose through central metabolism. The aim of this study was to further elucidate these metabolic changes by determining the activity and relative expression of key enzymes involved in glucose and glutamine metabolism. For both CHO and BHK cells, there was an increase in the activity of glutaminase, glutamate cehydrogenase and glutamine synthetase, suggesting an increased flux through the glutaminolysis pathway. The activity of glucose- 6-phosphate dehydrogenase and pyruvate carboxylase in CHO cells was also increased whilst lactate dehydrogenase activity remained unaltered, suggesting an increased flux to the pentose phosphate pathway and TCA cycle, respectively. The activity of these enzymes in BHK cells was unchanged. Quantitative RT-PCR showed that expression levels of glutaminase and pyruvate carboxylase were the same with and without G418, indicating that the differences in activities were likely due to post-translational modifications.",
keywords = "BHK, CHO, Enzymes, G418, Metabolic load, Metabolism, RT-PCR",
author = "Yallop, {C. A.} and N{\o}rby, {P. L.} and R. Jensen and H. Reinbach and I. Svendsen",
year = "2003",
doi = "10.1023/B:CYTO.0000009821.82741.8c",
language = "English",
volume = "42",
pages = "87--99",
journal = "Cytotechnology",
issn = "0920-9069",
publisher = "Springer",
number = "2",

}

RIS

TY - JOUR

T1 - Characterisation of G418-induced metabolic load in recombinant CHO and BHK cells

T2 - effect on the activity and expression of central metabolic enzymes

AU - Yallop, C. A.

AU - Nørby, P. L.

AU - Jensen, R.

AU - Reinbach, H.

AU - Svendsen, I.

PY - 2003

Y1 - 2003

N2 - In a previous article (Yallop and Svendsen 2001), recombinant CHO and BHK cell lines, expressing the human glucagon receptor and the gastric inhibitory peptide receptor, respectively, showed reduced growth rates and altered nutrient utilisation when grown with increasing concentrations of G418. This response was associated with an increased expression of the neor protein, while expression of the recombinant membrane receptors remained unaltered. The metabolic response was characterised in both cell lines by an increase in the specific rate of glutamine utilisation and in CHO cells by a decrease in the yield of lactate from glucose, suggesting a change in the flux of glucose through central metabolism. The aim of this study was to further elucidate these metabolic changes by determining the activity and relative expression of key enzymes involved in glucose and glutamine metabolism. For both CHO and BHK cells, there was an increase in the activity of glutaminase, glutamate cehydrogenase and glutamine synthetase, suggesting an increased flux through the glutaminolysis pathway. The activity of glucose- 6-phosphate dehydrogenase and pyruvate carboxylase in CHO cells was also increased whilst lactate dehydrogenase activity remained unaltered, suggesting an increased flux to the pentose phosphate pathway and TCA cycle, respectively. The activity of these enzymes in BHK cells was unchanged. Quantitative RT-PCR showed that expression levels of glutaminase and pyruvate carboxylase were the same with and without G418, indicating that the differences in activities were likely due to post-translational modifications.

AB - In a previous article (Yallop and Svendsen 2001), recombinant CHO and BHK cell lines, expressing the human glucagon receptor and the gastric inhibitory peptide receptor, respectively, showed reduced growth rates and altered nutrient utilisation when grown with increasing concentrations of G418. This response was associated with an increased expression of the neor protein, while expression of the recombinant membrane receptors remained unaltered. The metabolic response was characterised in both cell lines by an increase in the specific rate of glutamine utilisation and in CHO cells by a decrease in the yield of lactate from glucose, suggesting a change in the flux of glucose through central metabolism. The aim of this study was to further elucidate these metabolic changes by determining the activity and relative expression of key enzymes involved in glucose and glutamine metabolism. For both CHO and BHK cells, there was an increase in the activity of glutaminase, glutamate cehydrogenase and glutamine synthetase, suggesting an increased flux through the glutaminolysis pathway. The activity of glucose- 6-phosphate dehydrogenase and pyruvate carboxylase in CHO cells was also increased whilst lactate dehydrogenase activity remained unaltered, suggesting an increased flux to the pentose phosphate pathway and TCA cycle, respectively. The activity of these enzymes in BHK cells was unchanged. Quantitative RT-PCR showed that expression levels of glutaminase and pyruvate carboxylase were the same with and without G418, indicating that the differences in activities were likely due to post-translational modifications.

KW - BHK

KW - CHO

KW - Enzymes

KW - G418

KW - Metabolic load

KW - Metabolism

KW - RT-PCR

U2 - 10.1023/B:CYTO.0000009821.82741.8c

DO - 10.1023/B:CYTO.0000009821.82741.8c

M3 - Journal article

AN - SCOPUS:1642460583

VL - 42

SP - 87

EP - 99

JO - Cytotechnology

JF - Cytotechnology

SN - 0920-9069

IS - 2

ER -

ID: 210532777