7SKX

Ab initio structure of proteinase K from electron-counted MicroED data

7SKX の概要

• エントリーDOI: 10.2210/pdb7skx/pdb
• EMDBエントリー: 25184 25185
• 分子名称: Proteinase K, 5-amino-2,4,6-triiodobenzene-1,3-dicarboxylic acid, CALCIUM ION, ... (4 entities in total)
• 機能のキーワード: 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: 30248.84

構造登録者

Martynowycz, M.W.,Clabbers, M.T.B.,Hattne, J.,Gonen, T. (登録日: 2021-10-21, 公開日: 2022-06-08, 最終更新日: 2024-10-16)

主引用文献

Martynowycz, M.W.,Clabbers, M.T.B.,Hattne, J.,Gonen, T.
Ab initio phasing macromolecular structures using electron-counted MicroED data.
Nat.Methods, 19:724-729, 2022

PubMed Abstract: Structures of two globular proteins were determined ab initio using microcrystal electron diffraction (MicroED) data that were collected on a direct electron detector in counting mode. Microcrystals were identified using a scanning electron microscope (SEM) and thinned with a focused ion beam (FIB) to produce crystalline lamellae of ideal thickness. Continuous-rotation data were collected using an ultra-low exposure rate to enable electron counting in diffraction. For the first sample, triclinic lysozyme extending to a resolution of 0.87 Å, an ideal helical fragment of only three alanine residues provided initial phases. These phases were improved using density modification, allowing the entire atomic structure to be built automatically. A similar approach was successful on a second macromolecular sample, proteinase K, which is much larger and diffracted to a resolution of 1.5 Å. These results demonstrate that macromolecules can be determined to sub-ångström resolution by MicroED and that ab initio phasing can be successfully applied to counting data.

PubMed: 35637302
DOI: 10.1038/s41592-022-01485-4

実験手法

ELECTRON CRYSTALLOGRAPHY (1.5 Å)