8DN7

The crystal structure of the Pisum sativum Toc75 POTRA domains in complex with fab ax9

8DN7 の概要

• エントリーDOI: 10.2210/pdb8dn7/pdb
• 分子名称: Protein TOC75, chloroplastic, fabax9 Heavy Chain, fabax9 Light Chain, ... (4 entities in total)
• 機能のキーワード: membrane protein, chloroplast, membrane protein-immune system complex, membrane protein/immune system
• 由来する生物種: Pisum sativum (pea); 詳細
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 171179.82

構造登録者

Srinivasan, K.,Noinaj, N. (登録日: 2022-07-10, 公開日: 2023-05-17, 最終更新日: 2024-11-20)

主引用文献

Srinivasan, K.,Erramilli, S.K.,Chakravarthy, S.,Gonzalez, A.,Kossiakoff, A.,Noinaj, N.
Characterization of synthetic antigen binding fragments targeting Toc75 for the isolation of TOC in A. thaliana and P. sativum.
Structure, 31:595-606.e5, 2023

PubMed Abstract: Roughly 95% of the proteins that make up the chloroplast must be imported from the cytoplasm. The machinery responsible for the translocation of these cargo proteins is called the translocon at the outer membrane of chloroplast (TOC). The TOC core consists of three proteins, Toc34, Toc75, and Toc159; no high-resolution structure has been solved of fully assembled TOC from plants. Efforts toward determining the structure of the TOC have been hindered almost entirely by difficulties in producing sufficient yields for structural studies. In this study, we introduce an innovative method that utilizes synthetic antigen binding fragments (sABs) to isolate TOC directly from wild-type plant biomass including A. thaliana and P. sativum. Binding between the sABs and the POTRA domains was characterized by size-exclusion chromatography coupled with small-angle X-ray scattering (SEC-SAXS), X-ray crystallography, and isothermal titration calorimetry. We also demonstrate the isolation of the TOC from P. sativum, laying the framework for large-scale isolation and purification of TOC for functional and structural studies.

PubMed: 36977410
DOI: 10.1016/j.str.2023.03.002

実験手法

X-RAY DIFFRACTION (2 Å)