8THE

Cryo-EM structure of Pseudomonas aeruginosa TonB-dependent transporter PhuR in complex with synthetic antibody and heme

これはPDB形式変換不可エントリーです。

8THE の概要

• エントリーDOI: 10.2210/pdb8the/pdb
• EMDBエントリー: 41255
• 分子名称: Heme/hemoglobin uptake outer membrane receptor PhuR, Synthetic Antibody Heavy Chain, Synthetic Antibody Light Chain, ... (6 entities in total)
• 機能のキーワード: beta barrel, outer membrane protein, heme binding protein, heme transport, membrane protein
• 由来する生物種: Pseudomonas aeruginosa; 詳細
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 137743.51

構造登録者

Knejski, P.,Erramilli, S.K.,Kossiakoff, A.A. (登録日: 2023-07-15, 公開日: 2024-01-31, 最終更新日: 2024-11-13)

主引用文献

Knejski, P.P.,Erramilli, S.K.,Kossiakoff, A.A.
Chaperone-assisted cryo-EM structure of P. aeruginosa PhuR reveals molecular basis for heme binding.
Structure, 32:411-, 2024

PubMed Abstract: Pathogenic bacteria, such as Pseudomonas aeruginosa, depend on scavenging heme for the acquisition of iron, an essential nutrient. The TonB-dependent transporter (TBDT) PhuR is the major heme uptake protein in P. aeruginosa clinical isolates. However, a comprehensive understanding of heme recognition and TBDT transport mechanisms, especially PhuR, remains limited. In this study, we employed single-particle cryogenic electron microscopy (cryo-EM) and a phage display-generated synthetic antibody (sAB) as a fiducial marker to enable the determination of a high-resolution (2.5 Å) structure of PhuR with a bound heme. Notably, the structure reveals iron coordination by Y529 on a conserved extracellular loop, shedding light on the role of tyrosine in heme binding. Biochemical assays and negative-stain EM demonstrated that the sAB specifically targets the heme-bound state of PhuR. These findings provide insights into PhuR's heme binding and offer a template for developing conformation-specific sABs against outer membrane proteins (OMPs) for structure-function investigations.

PubMed: 38325368
DOI: 10.1016/j.str.2024.01.007

実験手法

ELECTRON MICROSCOPY (2.5 Å)