Lebreton, Sandrine and Graciet, Emmanuelle and Gontero, Brigitte (2003) Modulation, via Protein-Protein Interactions, of Glyceraldehyde-3-phosphate Dehydrogenase Activity through Redox Phosphoribulokinase Regulation. Journal of Biological Chemistry, 278 (14). pp. 12078-12084. ISSN 0021-9258

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Abstract

The activity of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) embedded in the phosphoribulokinase (PRK).GAPDH.CP12 complex was increased 2-3-fold by reducing agents. This occurred by interaction with PRK as the cysteinyl sulfhydryls (4 SH/subunit) of GAPDH within the complex were unchanged whatever the redox state of the complex. But isolated GAPDH was not activated. Alkylation plus mass spectrometry also showed that PRK had one disulfide bridge and three SH groups per monomer in the active oxidized complex. Reduction disrupted this disulfide bridge to give 2 more SH groups and a much more active enzyme. We assessed the kinetics and dynamics of the interactions between PRK and GAPDH/CP12 using biosensors to measure complex formation in real time. The apparent equilibrium binding constant for GAPDH/CP12 and PRK was 14 +/- 1.6 nm for oxidized PRK and 62 +/- 10 nm for reduced PRK. These interactions were neither pH- nor temperature-dependent. Thus, the dynamics of PRK.GAPDH.CP12 complex formation and GAPDH activity are modulated by the redox state of PRK.

Item Type:	Article
Additional Information:	This research was originally published in the Journal of Biological Chemistry: Lebreton S, Graciet E, Gontero B (2003) Modulation, via Protein-Protein Interactions, of Glyceraldehyde-3-phosphate Dehydrogenase Activity through Redox Phosphoribulokinase Regulation. Journal of Biological Chemistry, 278: 12078-12084
Keywords:	Modulation; Protein-Protein Interactions; Glyceraldehyde-3-phosphate Dehydrogenase Activity; Redox Phosphoribulokinase Regulation;
Academic Unit:	Faculty of Science and Engineering > Biology
Item ID:	7437
Identification Number:	https://doi.org/10.1074/jbc.M213096200
Depositing User:	Emanuelle Graciet
Date Deposited:	09 Sep 2016 15:14
Journal or Publication Title:	Journal of Biological Chemistry
Publisher:	American Society for Biochemistry and Molecular Biology
Refereed:	Yes
URI:	http://www.jbc.org
Use Licence:	This item is available under a Creative Commons Attribution Non Commercial Share Alike Licence (CC BY-NC-SA). Details of this licence are available here

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