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-Structure paper
タイトル | Bottom-up structural proteomics: cryoEM of protein complexes enriched from the cellular milieu. |
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ジャーナル・号・ページ | Nat Methods, Vol. 17, Issue 1, Page 79-85, Year 2020 |
掲載日 | 2019年11月25日 |
![]() | Chi-Min Ho / Xiaorun Li / Mason Lai / Thomas C Terwilliger / Josh R Beck / James Wohlschlegel / Daniel E Goldberg / Anthony W P Fitzpatrick / Z Hong Zhou / ![]() ![]() |
PubMed 要旨 | X-ray crystallography often requires non-native constructs involving mutations or truncations, and is challenged by membrane proteins and large multicomponent complexes. We present here a bottom-up ...X-ray crystallography often requires non-native constructs involving mutations or truncations, and is challenged by membrane proteins and large multicomponent complexes. We present here a bottom-up endogenous structural proteomics approach whereby near-atomic-resolution cryo electron microscopy (cryoEM) maps are reconstructed ab initio from unidentified protein complexes enriched directly from the endogenous cellular milieu, followed by identification and atomic modeling of the proteins. The proteins in each complex are identified using cryoID, a program we developed to identify proteins in ab initio cryoEM maps. As a proof of principle, we applied this approach to the malaria-causing parasite Plasmodium falciparum, an organism that has resisted conventional structural-biology approaches, to obtain atomic models of multiple protein complexes implicated in intraerythrocytic survival of the parasite. Our approach is broadly applicable for determining structures of undiscovered protein complexes enriched directly from endogenous sources. |
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手法 | EM (単粒子) |
解像度 | 3.2 Å |
構造データ | EMDB-20333, PDB-6pev: EMDB-20334, PDB-6pew: |
化合物 | ![]() ChemComp-ZN: |
由来 |
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![]() | METAL BINDING PROTEIN / aspartyl aminopeptidase / LIGASE / glutamine synthetase |