Understanding how glucose metabolism is finely regulated at molecular and cellular levels in the liver is critical for knowing its relationship to related pathologies, such as diabetes. In order to gain insight into the regulation of glucose metabolism, we studied the liver-expressed isoforms aldolase B and fructose-1,6-bisphosphatase-1 (FBPase-1), key enzymes in gluconeogenesis, analysing their cellular localization in hepatocytes under different metabolic conditions and their protein–protein interaction in vitro and in vivo. We observed that glucose, insulin, glucagon and adrenaline differentially modulate the intracellular distribution of aldolase B and FBPase-1. Interestingly, the in vitro protein–protein interaction analysis between aldolase B and FBPase-1 showed a specific and regulable interaction between them, whereas aldolase A (muscle isozyme) and FBPase-1 showed no interaction. The affinity of the aldolase B and FBPase-1 complex was modulated by intermediate metabolites, but only in the presence of K+. We observed a decreased association constant in the presence of adenosine monophosphate, fructose-2,6-bisphosphate, fructose-6-phosphate and inhibitory concentrations of fructose-1,6-bisphosphate. Conversely, the association constant of the complex increased in the presence of dihydroxyacetone phosphate (DHAP) and non-inhibitory concentrations of fructose-1,6-bisphosphate. Notably, in vivo FRET studies confirmed the interaction between aldolase B and FBPase-1. Also, the co-expression of aldolase B and FBPase-1 in cultured cells suggested that FBPase-1 guides the cellular localization of aldolase B. Our results provide further evidence that metabolic conditions modulate aldolase B and FBPase-1 activity at the cellular level through the regulation of their interaction, suggesting that their association confers a catalytic advantage for both enzymes.
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Research Article|
November 13 2015
A new level of regulation in gluconeogenesis: metabolic state modulates the intracellular localization of aldolase B and its interaction with liver fructose-1,6-bisphosphatase
Cristian A. Droppelmann;
Cristian A. Droppelmann
1
*Department of Biochemistry and Microbiology, Faculty of Sciences, Universidad Austral de Chile, Independencia 641,5110566, Valdivia, Chile
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Doris E. Sáez;
Doris E. Sáez
*Department of Biochemistry and Microbiology, Faculty of Sciences, Universidad Austral de Chile, Independencia 641,5110566, Valdivia, Chile
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Joel L. Asenjo;
Joel L. Asenjo
*Department of Biochemistry and Microbiology, Faculty of Sciences, Universidad Austral de Chile, Independencia 641,5110566, Valdivia, Chile
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Alejandro J. Yáñez;
Alejandro J. Yáñez
*Department of Biochemistry and Microbiology, Faculty of Sciences, Universidad Austral de Chile, Independencia 641,5110566, Valdivia, Chile
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Mar García-Rocha;
Mar García-Rocha
†Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute of Science and Technology, Baldiri Reixac 10, 08028 Barcelona, Spain
‡Department of Biochemistry and Molecular Biology, University of Barcelona, Diagonal 643, 08028, Barcelona, Spain
§CIBER of Diabetes and Metabolic Diseases (CIBERDEM), Instituto de Salud Carlos III, Monforte de Lemos 3-5, 28029 Madrid, Spain
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Ilona I. Concha;
Ilona I. Concha
*Department of Biochemistry and Microbiology, Faculty of Sciences, Universidad Austral de Chile, Independencia 641,5110566, Valdivia, Chile
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Manuel Grez;
Manuel Grez
║Institute for Tumor Biology and Experimental Therapy, Georg-Speyer Haus, Paul-Ehrlich-Straβe 42-44, D-60596, Frankfurt, Germany
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Joan J. Guinovart;
Joan J. Guinovart
†Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute of Science and Technology, Baldiri Reixac 10, 08028 Barcelona, Spain
‡Department of Biochemistry and Molecular Biology, University of Barcelona, Diagonal 643, 08028, Barcelona, Spain
§CIBER of Diabetes and Metabolic Diseases (CIBERDEM), Instituto de Salud Carlos III, Monforte de Lemos 3-5, 28029 Madrid, Spain
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Juan C. Slebe
Juan C. Slebe
2
*Department of Biochemistry and Microbiology, Faculty of Sciences, Universidad Austral de Chile, Independencia 641,5110566, Valdivia, Chile
2To whom correspondence should be addressed (email jslebe@uach.cl).
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Publisher: Portland Press Ltd
Received:
March 02 2015
Revision Received:
September 17 2015
Accepted:
September 25 2015
Accepted Manuscript online:
September 28 2015
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© 2015 Authors; published by Portland Press Limited
2015
Biochem J (2015) 472 (2): 225–237.
Article history
Received:
March 02 2015
Revision Received:
September 17 2015
Accepted:
September 25 2015
Accepted Manuscript online:
September 28 2015
Citation
Cristian A. Droppelmann, Doris E. Sáez, Joel L. Asenjo, Alejandro J. Yáñez, Mar García-Rocha, Ilona I. Concha, Manuel Grez, Joan J. Guinovart, Juan C. Slebe; A new level of regulation in gluconeogenesis: metabolic state modulates the intracellular localization of aldolase B and its interaction with liver fructose-1,6-bisphosphatase. Biochem J 1 December 2015; 472 (2): 225–237. doi: https://doi.org/10.1042/BJ20150269
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