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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Cox N, Charlier C, Vijayaraj R, De La Mare M, Barbe S, André I, Lippens G, and Montanier CY
- Subjects
- Amino Acids metabolism, Bacterial Proteins metabolism, Multiprotein Complexes metabolism, Protein Binding, Protein Stability, Proton Magnetic Resonance Spectroscopy, Streptococcus pneumoniae metabolism, Bacterial Proteins chemistry, Multiprotein Complexes chemistry
- Abstract
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., Competing Interests: Conflict of interest The authors (Neil Cox, Cyril Charlier, Ramadoss Vijayaraj, Marion De La Mare, Sophie Barbe, Isabelle André, Guy Lippens and Cédric Y. Montanier) declare that there is no conflict of interest., (Copyright © 2021 Elsevier Inc. All rights reserved.)
- Published
- 2022
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