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Microfluidic protein isolation and sample preparation for high-resolution cryo-EM

Schmidli, Claudio and Albiez, Stefan and Rima, Luca and Righetto, Ricardo and Mohammed, Inayatulla and Oliva, Paolo and Kovacik, Lubomir and Stahlberg, Henning and Braun, Thomas. (2019) Microfluidic protein isolation and sample preparation for high-resolution cryo-EM. Proceedings of the National Academy of Sciences of the United States of America, 116 (30). pp. 15007-15012.

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Official URL: https://edoc.unibas.ch/71491/

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Abstract

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.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Structural Biology & Biophysics > Structural Biology (Stahlberg)
UniBasel Contributors:Stahlberg, Henning
Item Type:Article, refereed
Article Subtype:Research Article
Publisher:National Academy of Sciences
ISSN:0027-8424
e-ISSN:1091-6490
Note:Publication type according to Uni Basel Research Database: Journal article
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Last Modified:17 Aug 2020 15:24
Deposited On:17 Aug 2020 14:51

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