+検索条件
-Structure paper
タイトル | Ab initio phasing macromolecular structures using electron-counted MicroED data. |
---|---|
ジャーナル・号・ページ | Nat Methods, Vol. 19, Issue 6, Page 724-729, Year 2022 |
掲載日 | 2022年5月30日 |
![]() | Michael W Martynowycz / Max T B Clabbers / Johan Hattne / Tamir Gonen / ![]() |
PubMed 要旨 | 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 ...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. |
![]() | ![]() ![]() ![]() |
手法 | EM (電子線結晶学) |
解像度 | 0.87 - 1.5 Å |
構造データ | EMDB-25184, PDB-7skw: EMDB-25185, PDB-7skx: |
化合物 | ![]() ChemComp-NO3: ![]() ChemComp-HOH: ![]() ChemComp-I3C: ![]() ChemComp-CA: |
由来 |
|
![]() | HYDROLASE |