8W9M

Cryo-EM structure of the cyanobacterial nitrate transporter NrtBCD in complex with ATP

8W9M の概要

• エントリーDOI: 10.2210/pdb8w9m/pdb
• EMDBエントリー: 37375
• 分子名称: Nitrate transport permease protein, Nitrate transport ATP-binding protein, ADENOSINE-5'-TRIPHOSPHATE, ... (5 entities in total)
• 機能のキーワード: atp-dependent transporter, abc transporter, nitrate/nitrite transporter, membrane protein
• 由来する生物種: Nostoc sp. PCC 7120 = FACHB-418; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 169752.39

構造登録者

Li, B.,Zhou, C.Z.,Chen, Y.X.,Jiang, Y.L. (登録日: 2023-09-05, 公開日: 2024-02-28, 最終更新日: 2024-03-20)

主引用文献

Li, B.,Wang, X.Q.,Li, Q.Y.,Xu, D.,Li, J.,Hou, W.T.,Chen, Y.,Jiang, Y.L.,Zhou, C.Z.
Allosteric regulation of nitrate transporter NRT via the signaling protein PII.
Proc.Natl.Acad.Sci.USA, 121:e2318320121-e2318320121, 2024

PubMed Abstract: Coordinated carbon and nitrogen metabolism is crucial for bacteria living in the fluctuating environments. Intracellular carbon and nitrogen homeostasis is maintained by a sophisticated network, in which the widespread signaling protein PII acts as a major regulatory hub. In cyanobacteria, PII was proposed to regulate the nitrate uptake by an ABC (ATP-binding cassette)-type nitrate transporter NrtABCD, in which the nucleotide-binding domain of NrtC is fused with a C-terminal regulatory domain (CRD). Here, we solved three cryoelectron microscopy structures of NrtBCD, bound to nitrate, ATP, and PII, respectively. Structural and biochemical analyses enable us to identify the key residues that form a hydrophobic and a hydrophilic cavity along the substrate translocation channel. The core structure of PII, but not the canonical T-loop, binds to NrtC and stabilizes the CRD, making it visible in the complex structure, narrows the substrate translocation channel in NrtB, and ultimately locks NrtBCD at an inhibited inward-facing conformation. Based on these results and previous reports, we propose a putative transport cycle driven by NrtABCD, which is allosterically inhibited by PII in response to the cellular level of 2-oxoglutarate. Our findings provide a distinct regulatory mechanism of ABC transporter via asymmetrically binding to a signaling protein.

PubMed: 38457518
DOI: 10.1073/pnas.2318320121

実験手法

ELECTRON MICROSCOPY (3.1 Å)