+検索条件
-Structure paper
タイトル | Cryo-EM of soft-landed β-galactosidase: Gas-phase and native structures are remarkably similar. |
---|---|
ジャーナル・号・ページ | Sci Adv, Vol. 10, Issue 7, Page eadl4628, Year 2024 |
掲載日 | 2024年2月16日 |
著者 | Tim K Esser / Jan Böhning / Alpcan Önür / Dinesh K Chinthapalli / Lukas Eriksson / Marko Grabarics / Paul Fremdling / Albert Konijnenberg / Alexander Makarov / Aurelien Botman / Christine Peter / Justin L P Benesch / Carol V Robinson / Joseph Gault / Lindsay Baker / Tanmay A M Bharat / Stephan Rauschenbach / |
PubMed 要旨 | Native mass spectrometry (MS) has become widely accepted in structural biology, providing information on stoichiometry, interactions, homogeneity, and shape of protein complexes. Yet, the fundamental ...Native mass spectrometry (MS) has become widely accepted in structural biology, providing information on stoichiometry, interactions, homogeneity, and shape of protein complexes. Yet, the fundamental assumption that proteins inside the mass spectrometer retain a structure faithful to native proteins in solution remains a matter of intense debate. Here, we reveal the gas-phase structure of β-galactosidase using single-particle cryo-electron microscopy (cryo-EM) down to 2.6-Å resolution, enabled by soft landing of mass-selected protein complexes onto cold transmission electron microscopy (TEM) grids followed by in situ ice coating. We find that large parts of the secondary and tertiary structure are retained from the solution. Dehydration-driven subunit reorientation leads to consistent compaction in the gas phase. By providing a direct link between high-resolution imaging and the capability to handle and select protein complexes that behave problematically in conventional sample preparation, the approach has the potential to expand the scope of both native mass spectrometry and cryo-EM. |
リンク | Sci Adv / PubMed:38354247 / PubMed Central |
手法 | EM (単粒子) |
解像度 | 2.3 - 2.6 Å |
構造データ | EMDB-18244, PDB-8q7y: EMDB-18245: Plunge-frozen (control) map of beta-galactosidase |
由来 |
|
キーワード | HYDROLASE / Lactase / Beta-galactosidase / cryo-EM / native MS |