Titre
Microfluidic protein isolation and sample preparation for high-resolution cryo-EM.
Type
article
Institution
Externe
Auteur(s)
Schmidli, C.
Auteure/Auteur
Albiez, S.
Auteure/Auteur
Rima, L.
Auteure/Auteur
Righetto, R.
Auteure/Auteur
Mohammed, I.
Auteure/Auteur
Oliva, P.
Auteure/Auteur
Kovacik, L.
Auteure/Auteur
Stahlberg, H.
Auteure/Auteur
Braun, T.
Auteure/Auteur
Liens vers les personnes
ISSN
1091-6490
Statut éditorial
Publié
Date de publication
2019-07-23
Volume
116
Numéro
30
Première page
15007
Dernière page/numéro d’article
15012
Peer-reviewed
Oui
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Publication Status: ppublish
Résumé
High-resolution structural information is essential to understand protein function. Protein-structure determination needs a considerable amount of protein, which can be challenging to produce, often involving harsh and lengthy procedures. In contrast, the several thousand to a few million protein particles required for structure determination by cryogenic electron microscopy (cryo-EM) can be provided by miniaturized systems. Here, we present a microfluidic method for the rapid isolation of a target protein and its direct preparation for cryo-EM. Less than 1 μL of cell lysate is required as starting material to solve the atomic structure of the untagged, endogenous human 20S proteasome. Our work paves the way for high-throughput structure determination of proteins from minimal amounts of cell lysate and opens more opportunities for the isolation of sensitive, endogenous protein complexes.
Sujets
PID Serval
serval:BIB_77C82DE8C722
PMID
Open Access
Oui
Date de création
2023-06-09T14:02:06.011Z
Date de création dans IRIS
2025-05-21T03:13:14Z