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-Structure paper
タイトル | Template-free detection and classification of membrane-bound complexes in cryo-electron tomograms. |
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ジャーナル・号・ページ | Nat Methods, Vol. 17, Issue 2, Page 209-216, Year 2020 |
掲載日 | 2020年1月6日 |
著者 | Antonio Martinez-Sanchez / Zdravko Kochovski / Ulrike Laugks / Johannes Meyer Zum Alten Borgloh / Saikat Chakraborty / Stefan Pfeffer / Wolfgang Baumeister / Vladan Lučić / |
PubMed 要旨 | With faithful sample preservation and direct imaging of fully hydrated biological material, cryo-electron tomography provides an accurate representation of molecular architecture of cells. However, ...With faithful sample preservation and direct imaging of fully hydrated biological material, cryo-electron tomography provides an accurate representation of molecular architecture of cells. However, detection and precise localization of macromolecular complexes within cellular environments is aggravated by the presence of many molecular species and molecular crowding. We developed a template-free image processing procedure for accurate tracing of complex networks of densities in cryo-electron tomograms, a comprehensive and automated detection of heterogeneous membrane-bound complexes and an unsupervised classification (PySeg). Applications to intact cells and isolated endoplasmic reticulum (ER) allowed us to detect and classify small protein complexes. This classification provided sufficiently homogeneous particle sets and initial references to allow subsequent de novo subtomogram averaging. Spatial distribution analysis showed that ER complexes have different localization patterns forming nanodomains. Therefore, this procedure allows a comprehensive detection and structural analysis of complexes in situ. |
リンク | Nat Methods / PubMed:31907446 |
手法 | EM (サブトモグラム平均) / EM (トモグラフィー) |
解像度 | 14.0 - 39.08 Å |
構造データ | EMDB-0074: EMDB-0075: EMDB-0084: EMDB-0085: EMDB-0086: EMDB-0087: EMDB-10432: EMDB-10433: EMDB-10434: EMDB-10435: EMDB-10436: EMDB-10437: EMDB-10439: EMDB-10449: EMDB-10450: EMDB-10451: |
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