8UY6

Aquaporin Z with ALFA tag and bound to nanobody

8UY6 の概要

• エントリーDOI: 10.2210/pdb8uy6/pdb
• EMDBエントリー: 42793
• 分子名称: Aquaporin Z, anti-ALFA nanobody, CARDIOLIPIN (3 entities in total)
• 機能のキーワード: aqpz, water channel, alfa tag, cardiolipin, membrane protein
• 由来する生物種: Escherichia coli; 詳細
• タンパク質・核酸の鎖数: 16
• 化学式量合計: 334193.71

構造登録者

Stover, L.,Bahramimoghaddam, H.,Wang, L.,Zhou, M.,Laganowsky, A. (登録日: 2023-11-13, 公開日: 2024-09-25, 最終更新日: 2024-10-16)

主引用文献

Stover, L.,Bahramimoghaddam, H.,Wang, L.,Schrecke, S.,Yadav, G.P.,Zhou, M.,Laganowsky, A.
Grafting the ALFA tag for structural studies of aquaporin Z.
J Struct Biol X, 9:100097-100097, 2024

PubMed Abstract: Aquaporin Z (AqpZ), a bacterial water channel, forms a tetrameric complex and, like many other membrane proteins, activity is regulated by lipids. Various methods have been developed to facilitate structure determination of membrane proteins, such as the use of antibodies. Here, we graft onto AqpZ the ALFA tag (AqpZ-ALFA), an alpha helical epitope, to make use of the high-affinity anti-ALFA nanobody (nB). Native mass spectrometry reveals the AqpZ-ALFA fusion forms a stable, 1:1 complex with nB. Single-particle cryogenic electron microscopy studies reveal the octameric (AqpZ-ALFA)(nB) complex forms a dimeric assembly and the structure was determined to 1.9 Å resolution. Dimerization of the octamer is mediated through stacking of the symmetrically bound nBs. Tube-like density is also observed, revealing a potential cardiolipin binding site. Grafting of the ALFA tag, or other epitope, along with binding and association of nBs to promote larger complexes will have applications in structural studies and protein engineering.

PubMed: 38361954
DOI: 10.1016/j.yjsbx.2024.100097

実験手法

ELECTRON MICROSCOPY (1.9 Å)