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Dynamics of the intrinsically disordered protein CP12 in its association with GAPDH in the green alga Chlamydomonas reinhardtii: a fuzzy complex

Elisabetta Mileo 1 Magali Lorenzi Jenny Erales 2 Sabrina Lignon 3 Carine Puppo 1 Nolwenn Le Breton 1 Emilien Etienne 1 Sylvain R.A. Marque 4 Bruno Guigliarelli 1 Brigitte Gontero 1 Valérie Belle 1
1 BIP - Bioénergétique et Ingénierie des Protéines
2 AFMB - Architecture et fonction des macromolécules biologiques
3 Plateforme Protéomique [Marseille]
IMM - Institut de Microbiologie de la Méditerranée
4 ICR - Institut de Chimie Radicalaire
Abstract : CP12 is a widespread regulatory protein of oxygenic photosynthetic organisms that contributes to the regulation of the Calvin cycle by forming a supra-molecular complex with at least two enzymes: glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and phosphoribulokinase (PRK). CP12 shares some similarities with intrinsically disordered proteins (IDPs) depending on its redox state. In this study, site-directed spin labeling (SDSL) combined with EPR spectroscopy was used to probe the dynamic behavior of CP12 from Chlamydomonas reinhardtii upon binding to GAPDH, the first step towards ternary complex formation. The two N-terminal cysteine residues were labeled using the classical approach while the tyrosine located at the C-terminal end of CP12 was modified following an original procedure. The results show that the label grafted at the C-terminal extremity is in the vicinity of the interaction site whereas the N-terminal region remains fully disordered upon binding to GAPDH. In conclusion, GAPDH-CP12 is a fuzzy complex, in which the N-terminal region of CP12 keeps a conformational freedom in the bound form. This fuzziness could be one of the keys to facilitate binding of PRK to CP12-GAPDH and to form the ternary supra-molecular complex.
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https://hal-amu.archives-ouvertes.fr/hal-01460463
Contributor : Didier Gigmes <>
Submitted on : Tuesday, February 7, 2017 - 4:43:20 PM
Last modification on : Tuesday, October 13, 2020 - 3:10:12 AM

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Elisabetta Mileo, Magali Lorenzi, Jenny Erales, Sabrina Lignon, Carine Puppo, et al.. Dynamics of the intrinsically disordered protein CP12 in its association with GAPDH in the green alga Chlamydomonas reinhardtii: a fuzzy complex. Molecular BioSystems, Royal Society of Chemistry, 2013, 9 (11), pp.2869--2876. ⟨10.1039/c3mb70190e⟩. ⟨hal-01460463⟩

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