9hqp
From Proteopedia
Cryo-EM structure of mouse TMEM16F-YFP purified and plunged using MISO (microfluidic isolation)
Structural highlights
FunctionANO6_MOUSE Small-conductance calcium-activated nonselective cation (SCAN) channel which acts as a regulator of phospholipid scrambling in platelets, osteoblasts and fetal thymocytes. Phospholipid scrambling results in surface exposure of phosphatidylserine which in platelets is essential to trigger the clotting system whereas in osteoblasts is essential for the deposition of hydroxyapatite during bone mineralization. Has calcium-dependent phospholipid scramblase activity; scrambles phosphatidylserine, phosphatidylcholine and galactosylceramide. Can generate outwardly rectifying chloride channel currents in airway epithelial cells and Jurkat T lymphocytes.[1] [2] [3] [4] [5] Publication Abstract from PubMedSingle-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 microg 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. MISO: microfluidic protein isolation enables single-particle cryo-EM structure determination from a single cell colony.,Eluru G, De Gieter S, Schenck S, Stroobants A, Shrestha B, Erbel P, Brunner JD, Efremov RG Nat Methods. 2025 Nov 13. doi: 10.1038/s41592-025-02894-x. PMID:41233542[6] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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