1. The covalent complex of Jo-In results from a long-lived, non-covalent intermediate state with near-native structure
- Author
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Neil, Cox, Cyril, Charlier, Ramadoss, Vijayaraj, Marion, De La Mare, Sophie, Barbe, Isabelle, André, Guy, Lippens, Cédric Y, Montanier, Toulouse Biotechnology Institute (TBI), Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Toulouse White Biotechnology (TWB), the Computing mesocenter of Region Midi-Pyrenees (CALMIP, Toulouse, France) ., Toulouse White Biotechnology (mIMHETIQ project, 2014-2016, and Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)
- Subjects
[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM] ,Protein Stability ,Proton Magnetic Resonance Spectroscopy ,Biophysics ,[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology ,Cell Biology ,Biochemistry ,Biomolecular welding tool ,[INFO.INFO-MO]Computer Science [cs]/Modeling and Simulation ,NMR ,Jo-in ,Streptococcus pneumoniae ,Bacterial Proteins ,Multiprotein Complexes ,[SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN] ,[SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology ,Molecular modelling ,Amino Acids ,[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM] ,Molecular Biology ,ComputingMilieux_MISCELLANEOUS ,Protein Binding - Abstract
International audience; Covalent protein complexes have been used to assemble enzymes in large scaffolds for biotechnology purposes. Although the catalytic mechanism of the covalent linking of such proteins is well known, the recognition and overall structural mechanisms driving the association are far less understood but could help further functional engineering of these complexes. Here, we study the Jo-In complex by NMR spectroscopy and molecular modelling. We characterize a transient non-covalent complex, with structural elements close to those in the final covalent complex. Using site specific mutagenesis, we further show that this non-covalent association is essential for the covalent complex to form
- Published
- 2022