Structure, dynamics and transferability of the metal-dependent polyhistidine tetramerization motif TetrHis for single-chain Fv antibodies.

Autor: Healey RD; IGF, University of Montpellier, CNRS, INSERM, Montpellier, France.; Sosei-Heptares, Steinmetz Building, Granta Park, Cambridge, CB21 6DG, UK., Couillaud L; IGF, University of Montpellier, CNRS, INSERM, Montpellier, France., Hoh F; Centre de Biologie Structurale, University of Montpellier, CNRS, INSERM, Montpellier, France., Mouhand A; Centre de Biologie Structurale, University of Montpellier, CNRS, INSERM, Montpellier, France., Fouillen A; IGF, University of Montpellier, CNRS, INSERM, Montpellier, France., Couvineau P; IGF, University of Montpellier, CNRS, INSERM, Montpellier, France., Granier S; IGF, University of Montpellier, CNRS, INSERM, Montpellier, France. sebastien.granier@igf.cnrs.fr., Leyrat C; IGF, University of Montpellier, CNRS, INSERM, Montpellier, France. cedric.leyrat@igf.cnrs.fr.
Jazyk: angličtina
Zdroj: Communications chemistry [Commun Chem] 2023 Jul 28; Vol. 6 (1), pp. 160. Date of Electronic Publication: 2023 Jul 28.
DOI: 10.1038/s42004-023-00962-x
Abstrakt: The polyhistidine (6XHis) motif is one of the most ubiquitous protein purification tags. The 6XHis motif enables the binding of tagged proteins to various metals, which can be advantageously used for purification with immobilized metal affinity chromatography. Despite its popularity, protein structures encompassing metal-bound 6XHis are rare. Here, we obtained a 2.5 Å resolution crystal structure of a single chain Fv antibody (scFv) bearing a C-terminal sortase motif, 6XHis and TwinStrep tags (LPETGHHHHHHWSHPQFEK[G 3 S] 3 WSHPQFEK). The structure, obtained in the presence of cobalt, reveals a unique tetramerization motif (TetrHis) stabilized by 8 Co 2+ ions. The TetrHis motif contains four 6 residues-long β-strands, and each metal center coordinates 3 to 5 residues, including all 6XHis histidines. By combining dynamic light scattering, small angle x-ray scattering and molecular dynamics simulations, We investigated the influence of Co 2+ on the conformational dynamics of scFv 2A2, observing an open/close equilibrium of the monomer and the formation of cobalt-stabilized tetramers. By using a similar scFv design, we demonstrate the transferability of the tetramerization property. This novel metal-dependent tetramerization motif might be used as a fiducial marker for cryoelectron microscopy of scFv complexes, or even provide a starting point for designing metal-loaded biomaterials.
(© 2023. The Author(s).)
Databáze: MEDLINE
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