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标题：Microfluidic protein isolation and sample preparation for high-resolution cryo-EM
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作者：Claudio Schmidli ; Stefan Albiez ; Luca Rima 等
期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2019
卷号：116
期号：30
页码：15007-15012
DOI：10.1073/pnas.1907214116
出版社：The National Academy of Sciences of the United States of America
摘要：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.
关键词：cryo-EM ; protein purification ; endogenous ; sample preparation ; microfluidics