8V9Y

X-ray crystal structure of nanobody JGFN4

8V9Y の概要

• エントリーDOI: 10.2210/pdb8v9y/pdb
• 分子名称: JGFN4 (2 entities in total)
• 機能のキーワード: antitoxin
• 由来する生物種: Camelidae
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 12935.39

構造登録者

Shi, K.,Moller, N.,Aihara, H. (登録日: 2023-12-10, 公開日: 2024-08-07, 最終更新日: 2024-10-23)

主引用文献

Gallant, J.P.,Hicks, D.,Shi, K.,Moeller, N.H.,Hoppe, B.,Lake, E.W.,Baehr, C.,Pravetoni, M.,Aihara, H.,LeBeau, A.M.
Identification and biophysical characterization of a novel domain-swapped camelid antibody specific for fentanyl.
J.Biol.Chem., 300:107502-107502, 2024

PubMed Abstract: Opioid use disorders (OUD) and overdoses are ever-evolving public health threats that continue to grow in incidence and prevalence in the United States and abroad. Current treatments consist of opioid receptor agonists and antagonists, which are safe and effective but still suffer from some limitations. Murine and humanized monoclonal antibodies (mAb) have emerged as an alternative and complementary strategy to reverse and prevent opioid-induced respiratory depression. To explore antibody applications beyond traditional heavy-light chain mAbs, we identified and biophysically characterized a novel single-domain antibody specific for fentanyl from a camelid variable-heavy-heavy (VHH) domain phage display library. Structural data suggested that VHH binding to fentanyl was facilitated by a unique domain-swapped dimerization mechanism, which accompanied a rearrangement of complementarity-determining region loops leading to the formation of a fentanyl-binding pocket. Structure-guided mutagenesis further identified an amino acid substitution that improved the affinity and relaxed the requirement for dimerization of the VHH in fentanyl binding. Our studies demonstrate VHH engagement of an opioid and inform on how to further engineer a VHH for enhanced stability and efficacy, laying the groundwork for exploring the in vivo applications of VHH-based biologics against OUD and overdose.

PubMed: 38945452
DOI: 10.1016/j.jbc.2024.107502

実験手法

X-RAY DIFFRACTION (1.76 Å)