6R70

Endogeneous native human 20S proteasome

Summary for 6R70

• Entry DOI: 10.2210/pdb6r70/pdb
• EMDB information: 4628 4738
• Descriptor: Proteasome subunit beta type-4, Proteasome subunit beta type-3, Proteasome subunit beta type-2, ... (14 entities in total)
• Functional Keywords: native, human, 20s proteasome, proteasome, cryo-em, microfluidic, cryowriter, protein isolation, endogeneous, hydrolase
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 28
• Total formula weight: 693576.33

Authors

Schmidli, C.,Albiez, S.,Rima, L.,Righetto, R.,Mohammed, I.,Oliva, P.,Kovacik, L.,Stahlberg, H.,Braun, T. (deposition date: 2019-03-28, release date: 2019-07-03, Last modification date: 2024-05-22)

Primary citation

Schmidli, C.,Albiez, S.,Rima, L.,Righetto, R.,Mohammed, I.,Oliva, P.,Kovacik, L.,Stahlberg, H.,Braun, T.
Microfluidic protein isolation and sample preparation for high-resolution cryo-EM.
Proc.Natl.Acad.Sci.USA, 116:15007-15012, 2019

PubMed 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.

PubMed: 31292253
DOI: 10.1073/pnas.1907214116

Experimental method

ELECTRON MICROSCOPY (3.5 Å)