title: FRET analysis of CP12 structural interplay by GAPDH and PRK.
abstract: CP12 is an intrinsically disordered protein playing a key role in the regulation of the Benson-Calvin cycle. Due to the high intrinsic flexibility of CP12, it is essential to consider its structural modulation induced upon binding to the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and phosphoribulokinase (PRK) enzymes. Here, we report for the first time detailed structural modulation about the wild-type CP12 and its site-specific N-terminal and C-terminal disulfide bridge mutants upon interaction with GAPDH and PRK by Förster resonance energy transfer (FRET). Our results indicate an increase in CP12 compactness when the complex is formed with GAPDH or PRK. In addition, the distributions in FRET histograms show the elasticity and conformational flexibility of CP12 in all supra molecular complexes. Contrarily to previous beliefs, our FRET results importantly reveal that both N-terminal and C-terminal site-specific CP12 mutants are able to form the monomeric (GAPDH-CP12-PRK) complex.
authors: Aix Marseille Université, CNRS, Centrale Marseille, Institut Fresnel, UMR 7249, 13013 Marseille, France. Electronic address: satish.moparthi@fresnel.fr.
authors: Satish Babu
authors: SB
authors: Moparthi
authors: Aix Marseille Université, CNRS, UMR 7281 Laboratoire de Bioénergétique et Ingénierie des Protéines, 13402 Marseille Cedex 20, France.
authors: Gabriel
authors: G
authors: Thieulin-Pardo
authors: Aix Marseille Université, CNRS, Centrale Marseille, Institut Fresnel, UMR 7249, 13013 Marseille, France.
authors: Juan
authors: J
authors: de Torres
authors: Aix Marseille Université, CNRS, Centrale Marseille, Institut Fresnel, UMR 7249, 13013 Marseille, France.
authors: Petru
authors: P
authors: Ghenuche
authors: Aix Marseille Université, CNRS, UMR 7281 Laboratoire de Bioénergétique et Ingénierie des Protéines, 13402 Marseille Cedex 20, France.
authors: Brigitte
authors: B
authors: Gontero
authors: Aix Marseille Université, CNRS, Centrale Marseille, Institut Fresnel, UMR 7249, 13013 Marseille, France.
authors: Jérôme
authors: J
authors: Wenger
mesh_terms: D016825:Chlamydomonas reinhardtii / Q000737:chemistry / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry / Q000378:metabolism*; D010788:Photosynthesis; D010940:Plant Proteins / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D060066:Protein Interaction Maps
chemical_list: D010940:Plant Proteins; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D017853:Phosphotransferases (Alcohol Group Acceptor); C022390:phosphoribulokinase
keywords: CP12; Conformational modulation; GAPDH; Intrinsic disorder proteins; PRK; Single molecule FRET