Effects of overexpression of the liver subunit of 6-phosphofructo-1-kinase on the metabolism of a cultured mammalian cell line

Urbano, Ana M.; Gillham, Helen; Groner, Yoram; Brindle, Kevin M.

doi:10.1042/bj3520921

Cited by 15 publications

(9 citation statements)

References 43 publications

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“…We first restate that phosphofructokinase in not rate-limiting when operating in situ. This has been shown experimentally many times [73][74][75][76][77][78][79][80][81][82] as well as being consistent with theory. This cannot be easily disputed.…”

Section: The Phosphofructokinase Paradoxsupporting

confidence: 87%

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Control and regulation of pathways via negative feedback

Sauro¹

2017

J. R. Soc. Interface.

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The biochemical networks found in living organisms include a huge variety of control mechanisms at multiple levels of organization. While the mechanistic and molecular details of many of these control mechanisms are understood, their exact role in driving cellular behaviour is not. For example, yeast glycolysis has been studied for almost 80 years but it is only recently that we have come to understand the systemic role of the multitude of feedback and feed-forward controls that exist in this pathway. In this article, control theory is discussed as an approach to dissect the control logic of complex pathways. One of the key issues is distinguishing between the terms control and regulation and how these concepts are applied to regulated enzymes such as phosphofructokinase. In doing so, one of the paradoxes in metabolic regulation can be resolved where enzymes such as phosphofructokinase have little control but, nevertheless, possess significant regulatory influence.

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Section: The Phosphofructokinase Paradoxsupporting

confidence: 87%

“…This has been shown experimentally many times [73][74][75][76][77][78][79][80][81][82] as well as being consistent with theory. This cannot be easily disputed.…”

Section: The Phosphofructokinase Paradoxsupporting

confidence: 67%

Control and regulation of pathways via negative feedback

Sauro¹

2017

J. R. Soc. Interface.

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“…In addition to S. cerevisiae , overexpression of glycolytic enzymes in other organisms such as E. coli [ 15 , 16 ], lactobacteria [ 17 ], tomato [ 21 ], potato [ 22 ], and hamster ovary cells [ 19 ] has been accomplished, although without increasing flux ( Table 1 ). It is somewhat surprising to note that in the glycolytic enzyme overexpression experiments, the strong inhibitory effect of G6P (or Tre6P in S. cerevisiae ), and citrate on HK and PFK-1, respectively, have been neglected.…”

Section: In Vivo Overexpression Experiments Of Enzymesmentioning

confidence: 99%

Metabolic Control Analysis: A Tool for Designing Strategies to Manipulate Metabolic Pathways

Moreno‐Sánchez

Saavedra

Rodrı́guez-Enrı́quez

et al. 2008

BioMed Research International

166

164

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The traditional experimental approaches used for changing the flux or the concentration of a particular metabolite of a metabolic pathway have been mostly based on the inhibition or over-expression of the presumed rate-limiting step. However, the attempts to manipulate a metabolic pathway by following such approach have proved to be unsuccessful. Metabolic Control Analysis (MCA) establishes how to determine, quantitatively, the degree of control that a given enzyme exerts on flux and on the concentration of metabolites, thus substituting the intuitive, qualitative concept of rate limiting step. Moreover, MCA helps to understand (i) the underlying mechanisms by which a given enzyme exerts high or low control and (ii) why the control of the pathway is shared by several pathway enzymes and transporters. By applying MCA it is possible to identify the steps that should be modified to achieve a successful alteration of flux or metabolite concentration in pathways of biotechnological (e.g., large scale metabolite production) or clinical relevance (e.g., drug therapy). The different MCA experimental approaches developed for the determination of the flux-control distribution in several pathways are described. Full understanding of the pathway properties when is working under a variety of conditions can help to attain a successful manipulation of flux and metabolite concentration.

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“…The biochemical significance of a change in enzyme expression depends in complex fashion upon the chemical properties of its reaction mechanism and reaction network. For example, using in vitro systems, a four-to-five-fold increase in phosphofructokinase expression has no effect on glycolytic flux or cell growth 44, 45. However, similar changes in pyruvate kinase expression dramatically alter carbon flux and cell growth 46, 47.…”

Section: Discussionmentioning

confidence: 99%

Leukocyte Pyruvate Kinase Expression is Reduced in Normal Human Pregnancy but not in Pre‐eclampsia

Madsen–Bouterse

Romero

et al. 2010

American J Rep Immunol

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Problem Emerging evidence suggests that metabolism influences immune cell signaling and immunoregulation. To examine the immunoregulatory role of glycolysis in pregnancy, we evaluated the properties of pyruvate kinase in leukocytes from non-pregnant women and those with normal pregnancy and pre-eclampsia. Method of study We evaluated pyruvate kinase expression in lymphocytes and neutrophils from non-pregnant, pregnant, and pre-eclampsia patients using fluorescence microscopy and flow cytometry. Leukocyte pyruvate kinase activity and pyruvate concentrations were also evaluated. To study pyruvate’s effect on signaling, we labeled Jurkat T cells with Ca2+ dyes and measured cell responses in the presence of agents influencing intracellular pyruvate. Results The expression of pyruvate kinase is reduced in lymphocytes and neutrophils from normal pregnant women in comparison to those of non-pregnant women and pre-eclampsia patients. Similarly, the activity of pyruvate kinase and the intracellular pyruvate concentration are reduced in leukocytes of normal pregnant women in comparison to non-pregnant women and women with pre-eclampsia. Using Jurkat cells as a model of leukocyte signaling, we have shown that perturbations of intracellular pyruvate influence Ca2+ signals. Conclusion Normal pregnancy is characterized by reduced pyruvate kinase expression within lymphocytes and neutrophils. We speculate that reduced pyruvate kinase expression modifies immune cell responses due to reduced pyruvate concentrations.

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Effects of overexpression of the liver subunit of 6-phosphofructo-1-kinase on the metabolism of a cultured mammalian cell line

Cited by 15 publications

References 43 publications

Control and regulation of pathways via negative feedback

Control and regulation of pathways via negative feedback

Metabolic Control Analysis: A Tool for Designing Strategies to Manipulate Metabolic Pathways

Leukocyte Pyruvate Kinase Expression is Reduced in Normal Human Pregnancy but not in Pre‐eclampsia

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