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TitleClassification of Single Particles from Human Cell Extract Reveals Distinct Structures.
Journal, issue, pagesCell Rep, Vol. 24, Issue 1, Page 259-268.e3, Year 2018
Publish dateJul 3, 2018
AuthorsEric J Verbeke / Anna L Mallam / Kevin Drew / Edward M Marcotte / David W Taylor /
PubMed AbstractMulti-protein complexes are necessary for nearly all cellular processes, and understanding their structure is required for elucidating their function. Current high-resolution strategies in structural ...Multi-protein complexes are necessary for nearly all cellular processes, and understanding their structure is required for elucidating their function. Current high-resolution strategies in structural biology are effective but lag behind other fields (e.g., genomics and proteomics) due to their reliance on purified samples rather than heterogeneous mixtures. Here, we present a method combining single-particle analysis by electron microscopy with protein identification by mass spectrometry to structurally characterize macromolecular complexes from human cell extract. We identify HSP60 through two-dimensional classification and obtain three-dimensional structures of native proteasomes directly from ab initio classification of a heterogeneous mixture of protein complexes. In addition, we reveal an ∼1-MDa-size structure of unknown composition and reference our proteomics data to suggest possible identities. Our study shows the power of using a shotgun approach to electron microscopy (shotgun EM) when coupled with mass spectrometry as a tool to uncover the structures of macromolecular machines.
External linksCell Rep / PubMed:29972786 / PubMed Central
MethodsEM (single particle)
Resolution21.0 - 31.0 Å
Structure data

EMDB-7946:
Classification of Single Particles from Human Cell Extract Reveals Distinct Structures
Method: EM (single particle) / Resolution: 21.0 Å

EMDB-7947:
Classification of Single Particles from Human Cell Extract Reveals Distinct Structures
Method: EM (single particle) / Resolution: 31.0 Å

Source
  • Homo sapiens (human)

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