The covalent complex of Jo-In results from a long-lived, non-covalent intermediate state with near-native structure
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...
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Published in | Biochemical and biophysical research communications Vol. 589; pp. 223 - 228 |
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Main Authors | , , , , , , , |
Format | Journal Article |
Language | English |
Published |
United States
Elsevier Inc
22.01.2022
Elsevier |
Subjects | |
Online Access | Get full text |
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Summary: | 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.
•The covalent bond formation is not essential for Jo-In interaction.•A specific couple of amino acids at the Jo/In interface is engaged in the recognition and the stabilisation of the complex.•A transient complex formation is required for covalent bond formation between Jo and In. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 0006-291X 1090-2104 |
DOI: | 10.1016/j.bbrc.2021.12.028 |