6PKP

MicroED structure of proteinase K from a platinum-coated, polished, single lamella at 1.91A resolution (#10)

6PKP の概要

• エントリーDOI: 10.2210/pdb6pkp/pdb
• EMDBエントリー: 20356 20357 20358 20359 20360 20361 20362 20363 20364 20365 20366
• 分子名称: Proteinase K (2 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: 28930.78

構造登録者

Martynowycz, M.W.,Zhao, W.,Hattne, J.,Jensen, G.J.,Gonen, T. (登録日: 2019-06-29, 公開日: 2019-09-04, 最終更新日: 2019-12-18)

主引用文献

Martynowycz, M.W.,Zhao, W.,Hattne, J.,Jensen, G.J.,Gonen, T.
Qualitative Analyses of Polishing and Precoating FIB Milled Crystals for MicroED.
Structure, 27:1594-, 2019

PubMed Abstract: Microcrystal electron diffraction (MicroED) leverages the strong interaction between matter and electrons to determine protein structures from vanishingly small crystals. This strong interaction limits the thickness of crystals that can be investigated by MicroED, mainly due to absorption. Recent studies have demonstrated that focused ion-beam (FIB) milling can thin crystals into ideal-sized lamellae; however, it is not clear how to best apply FIB milling for MicroED. Here, the effects of polishing the lamellae, whereby the last few nanometers are milled away using a low-current gallium beam, are explored in both the platinum-precoated and uncoated samples. Our results suggest that precoating samples with a thin layer of platinum followed by polishing the crystal surfaces prior to data collection consistently led to superior results as indicated by higher signal-to-noise ratio, higher resolution, and better refinement statistics. This study lays the foundation for routine and reproducible methodology for sample preparation in MicroED.

PubMed: 31422911
DOI: 10.1016/j.str.2019.07.004

実験手法

ELECTRON CRYSTALLOGRAPHY (1.91 Å)