6PU4

MicroED structure of proteinase K recorded on Falcon III

6PU4 の概要

• エントリーDOI: 10.2210/pdb6pu4/pdb
• EMDBエントリー: 20475 20476
• 分子名称: Proteinase K, CALCIUM ION (3 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: 29091.10

構造登録者

Hattne, J.,Martynowycz, M.W.,Penzcek, P.A.,Gonen, T. (登録日: 2019-07-17, 公開日: 2019-08-28, 最終更新日: 2024-10-30)

主引用文献

Hattne, J.,Martynowycz, M.W.,Penczek, P.A.,Gonen, T.
MicroED with the Falcon III direct electron detector.
Iucrj, 6:921-926, 2019

PubMed Abstract: Microcrystal electron diffraction (MicroED) combines crystallography and electron cryo-microscopy (cryo-EM) into a method that is applicable to high-resolution structure determination. In MicroED, nanosized crystals, which are often intractable using other techniques, are probed by high-energy electrons in a transmission electron microscope. Diffraction data are recorded by a camera in movie mode: the nanocrystal is continuously rotated in the beam, thus creating a sequence of frames that constitute a movie with respect to the rotation angle. Until now, diffraction-optimized cameras have mostly been used for MicroED. Here, the use of a direct electron detector that was designed for imaging is reported. It is demonstrated that data can be collected more rapidly using the Falcon III for MicroED and with markedly lower exposure than has previously been reported. The Falcon III was operated at 40 frames per second and complete data sets reaching atomic resolution were recorded in minutes. The resulting density maps to 2.1 Å resolution of the serine protease proteinase K showed no visible signs of radiation damage. It is thus demonstrated that dedicated diffraction-optimized detectors are not required for MicroED, as shown by the fact that the very same cameras that are used for imaging applications in electron microscopy, such as single-particle cryo-EM, can also be used effectively for diffraction measurements.

PubMed: 31576224
DOI: 10.1107/S2052252519010583

実験手法

ELECTRON CRYSTALLOGRAPHY (2.1 Å)