8CGE

The crystal structure of a cobalt-bound scFv reveals a Tetrameric polyHistidine motif (TetrHis)

8CGE の概要

• エントリーDOI: 10.2210/pdb8cge/pdb
• 分子名称: Single Chain Fv 2A2, COBALT (II) ION, SULFATE ION, ... (5 entities in total)
• 機能のキーワード: single chain fragment variable antibody, tetrameric polyhistidine tetrhis motif, cobalt-bound, metal binding protein
• 由来する生物種: synthetic construct
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 66521.76

構造登録者

Healey, R.D.,Hoh, F.,Granier, S.,Leyrat, C. (登録日: 2023-02-03, 公開日: 2023-08-09, 最終更新日: 2024-11-13)

主引用文献

Healey, R.D.,Couillaud, L.,Hoh, F.,Mouhand, A.,Fouillen, A.,Couvineau, P.,Granier, S.,Leyrat, C.
Structure, dynamics and transferability of the metal-dependent polyhistidine tetramerization motif TetrHis for single-chain Fv antibodies.
Commun Chem, 6:160-160, 2023

PubMed Abstract: 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[GS]WSHPQFEK). The structure, obtained in the presence of cobalt, reveals a unique tetramerization motif (TetrHis) stabilized by 8 Co 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 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.

PubMed: 37507458
DOI: 10.1038/s42004-023-00962-x

実験手法

X-RAY DIFFRACTION (2.5 Å)