9HQP
Cryo-EM structure of mouse TMEM16F-YFP purified and plunged using MISO (microfluidic isolation)
Summary for 9HQP
| Entry DOI | 10.2210/pdb9hqp/pdb |
| EMDB information | 52346 |
| Descriptor | Anoctamin-6,Yellow Fluorescent Protein, CALCIUM ION (2 entities in total) |
| Functional Keywords | lipid scramblase, membrane protein |
| Biological source | Mus musculus (house mouse) More |
| Total number of polymer chains | 2 |
| Total formula weight | 281414.00 |
| Authors | De Gieter, S.,Eluru, G.,Schenck, S.,Stroobants, A.,Efremov, R.G.,Brunner, J.D. (deposition date: 2024-12-16, release date: 2025-11-26) |
| 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, 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 (3.51 Å) |
Structure validation
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