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

Type of publication Peer-reviewed
Publikationsform Original article (peer-reviewed)
Author Schmidli Claudio, Albiez Stefan, Rima Luca, Righetto Ricardo, Mohammed Inayatulla, Oliva Paolo, Kovacik Lubomir, Stahlberg Henning, Braun Thomas,
Project Fast protein-complex isolation, sample preparation and data processing for high-resolution structural analysis and visual proteomics
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Original article (peer-reviewed)

Journal Proceedings of the National Academy of Sciences
Page(s) 201907214 - 201907214
Title of proceedings Proceedings of the National Academy of Sciences
DOI 10.1073/pnas.1907214116

Open Access

URL http://doi.org/10.1073/pnas.1907214116
Type of Open Access Publisher (Gold Open Access)

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.
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