9HQP

Cryo-EM structure of mouse TMEM16F-YFP purified and plunged using MISO (microfluidic isolation)

9HQP の概要

• エントリーDOI: 10.2210/pdb9hqp/pdb
• EMDBエントリー: 52346
• 分子名称: Anoctamin-6,Yellow Fluorescent Protein, CALCIUM ION (2 entities in total)
• 機能のキーワード: lipid scramblase, membrane protein
• 由来する生物種: Mus musculus (house mouse); 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 281414.00

構造登録者

De Gieter, S.,Eluru, G.,Schenck, S.,Stroobants, A.,Efremov, R.G.,Brunner, J.D. (登録日: 2024-12-16, 公開日: 2025-11-26)

主引用文献

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

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: 41233542
DOI: 10.1038/s41592-025-02894-x

実験手法

ELECTRON MICROSCOPY (3.51 Å)