9HQO
Cryo-EM structure of bovine TMEM206-YFP purified and plunged using MISO (micro-purification)
Summary for 9HQO
| Entry DOI | 10.2210/pdb9hqo/pdb |
| EMDB information | 52345 |
| Descriptor | Proton-activated chloride channel (1 entity in total) |
| Functional Keywords | ph-gated chloride channel, membrane protein |
| Biological source | Bos taurus (domestic cattle) |
| Total number of polymer chains | 3 |
| Total formula weight | 91127.64 |
| Authors | De Gieter, S.,Eluru, G.,Schenck, S.,Stroobants, A.,Efremov, R.G.,Brunner, J.D. (deposition date: 2024-12-16, release date: 2025-11-05, Last modification date: 2025-12-24) |
| Primary citation | Eluru, G.,De Gieter, S.,Schenck, S.,Stroobants, A.,Shrestha, B.,Erbel, P.,Brunner, J.D.,Efremov, R.G. MISO: microfluidic protein isolation enables single-particle cryo-EM structure determination from a single cell colony. Nat.Methods, 22:2563-2573, 2025 Cited by PubMed Abstract: Single-particle cryogenic electron microscopy (cryo-EM) enables reconstruction of atomic-resolution 3D maps of proteins by visualizing thousands to millions of purified protein particles embedded in vitreous ice. This corresponds to picograms of purified protein, which can potentially be isolated from a few thousand cells. Hence, cryo-EM holds the potential of a very sensitive analytical method for delivering high-resolution protein structure as a readout. In practice, millions of times more starting biological material is required to prepare cryo-EM grids. Here we show that using a micro isolation (MISO) method, which combines microfluidics-based protein purification with cryo-EM grid preparation, cryo-EM structures of soluble bacterial and eukaryotic membrane proteins can be solved starting from less than 1 µg of a target protein and progressing from cells to cryo-EM grids within a few hours. This scales down the amount of starting biological material hundreds to thousands of times, opening possibilities for the structural characterization of hitherto inaccessible proteins. PubMed: 41233542DOI: 10.1038/s41592-025-02894-x PDB entries with the same primary citation |
| Experimental method | ELECTRON MICROSCOPY (2.97 Å) |
Structure validation
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