6N4U

MicroED structure of Proteinase K at 2.75A resolution from a single milled crystal.

6N4U の概要

• エントリーDOI: 10.2210/pdb6n4u/pdb
• EMDBエントリー: 0343
• 分子名称: Proteinase K, CALCIUM ION, SULFATE ION, ... (4 entities in total)
• 機能のキーワード: broad-spectrum serum proteinase, hydrolase
• 由来する生物種: Engyodontium album
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 29135.01

構造登録者

Martynowycz, M.W.,Zhao, W.,Hattne, J.,Jensen, G.J.,Gonen, T. (登録日: 2018-11-20, 公開日: 2019-02-06, 最終更新日: 2023-10-11)

主引用文献

Martynowycz, M.W.,Zhao, W.,Hattne, J.,Jensen, G.J.,Gonen, T.
Collection of Continuous Rotation MicroED Data from Ion Beam-Milled Crystals of Any Size.
Structure, 27:545-548.e2, 2019

PubMed Abstract: Microcrystal electron diffraction (MicroED) allows for macromolecular structure solution from nanocrystals. To create crystals of suitable size for MicroED data collection, sample preparation typically involves sonication or pipetting a slurry of crystals from a crystallization drop. The resultant crystal fragments are fragile and the quality of the data that can be obtained from them is sensitive to subsequent sample preparation for cryoelectron microscopy as interactions in the water-air interface can damage crystals during blotting. Here, we demonstrate the use of a focused ion beam to generate lamellae of macromolecular protein crystals for continuous rotation MicroED that are of ideal thickness, easy to locate, and require no blotting optimization. In this manner, crystals of nearly any size may be scooped and milled to desired dimensions prior to data collection, thus streamlining the methodology for sample preparation for MicroED.

PubMed: 30661853
DOI: 10.1016/j.str.2018.12.003

実験手法

ELECTRON CRYSTALLOGRAPHY (2.75 Å)