7JSY

Proteinase K soaked with I3C determined by MicroED from a single milled microcrystal

7JSY の概要

• エントリーDOI: 10.2210/pdb7jsy/pdb
• EMDBエントリー: 22463
• 分子名称: Proteinase K, 5-amino-2,4,6-triiodobenzene-1,3-dicarboxylic acid (3 entities in total)
• 機能のキーワード: proteinase k, hydrolase
• 由来する生物種: Parengyodontium album (Engyodontium album, Tritirachium album)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 31166.12

構造登録者

Martynowycz, M.W.,Gonen, T. (登録日: 2020-08-16, 公開日: 2020-10-14, 最終更新日: 2024-11-13)

主引用文献

Martynowycz, M.W.,Gonen, T.
Ligand Incorporation into Protein Microcrystals for MicroED by On-Grid Soaking.
Structure, 29:88-, 2021

PubMed Abstract: A high throughout method for soaking ligands into protein microcrystals on TEM grids is presented. Every crystal on the grid is soaked simultaneously using only standard cryoelectron microscopy vitrification equipment. The method is demonstrated using proteinase K microcrystals soaked with the 5-amino-2,4,6-triodoisophthalic acid (I3C) magic triangle. A soaked microcrystal is milled to a thickness of approximately 200 nm using a focused ion beam, and MicroED data are collected. A high-resolution structure of the protein with four ligands at high occupancy is determined. Both the number of ligands bound and their occupancy is higher using on-grid soaking of microcrystals compared with much larger crystals treated similarly and investigated by X-ray crystallography. These results indicate that on-grid soaking ligands into microcrystals results in efficient uptake of ligands into protein microcrystals.

PubMed: 33007196
DOI: 10.1016/j.str.2020.09.003

実験手法

ELECTRON CRYSTALLOGRAPHY (1.78 Å)