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TitleToward community-driven visual proteomics with large-scale cryo-electron tomography of Chlamydomonas reinhardtii.
Journal, issue, pagesMol Cell, Vol. 86, Issue 1, Page 213-230.e7, Year 2026
Publish dateJan 8, 2026
AuthorsRon Kelley / Sagar Khavnekar / Ricardo D Righetto / Jessica Heebner / Martin Obr / Xianjun Zhang / Saikat Chakraborty / Grigory Tagiltsev / Alicia K Michael / Sofie van Dorst / Florent Waltz / Caitlyn L McCafferty / Lorenz Lamm / Simon Zufferey / Philippe Van der Stappen / Hugo van den Hoek / Wojciech Wietrzynski / Pavol Harar / William Wan / John A G Briggs / Jürgen M Plitzko / Benjamin D Engel / Abhay Kotecha /
PubMed AbstractIn situ cryo-electron tomography (cryo-ET) has emerged as the method of choice to investigate the structures of biomolecules in their native context. However, challenges remain for the efficient ...In situ cryo-electron tomography (cryo-ET) has emerged as the method of choice to investigate the structures of biomolecules in their native context. However, challenges remain for the efficient production and sharing of large-scale cryo-ET datasets. Here, we combined cryogenic plasma-based focused ion beam (cryo-PFIB) milling with recent advances in cryo-ET acquisition and processing to generate a dataset of 1,829 annotated tomograms of the green alga Chlamydomonas reinhardtii, which we provide as a community resource to drive method development and inspire biological discovery. To assay data quality, we performed subtomogram averaging of both soluble and membrane-bound complexes ranging in size from >3 MDa to ∼200 kDa, including 80S ribosomes, Rubisco, nucleosomes, microtubules, clathrin, photosystem II, and mitochondrial ATP synthase. The majority of these density maps reached sub-nanometer resolution, demonstrating the potential of this C. reinhardtii dataset as well as the promise of modern cryo-ET workflows and open data sharing to empower visual proteomics.
External linksMol Cell / PubMed:41421337
MethodsEM (subtomogram averaging)
Resolution4.0 - 18.8 Å
Structure data

EMDB-19906: In situ nucleosome subtomogram average from Chlamydomonas reinhardtii
Method: EM (subtomogram averaging) / Resolution: 9.65 Å

EMDB-51731: In situ PSII subtomogram average from Chlamydomonas reinhardtii
Method: EM (subtomogram averaging) / Resolution: 18.8 Å

EMDB-51789: In situ clathrin subtomogram average from Chlamydomonas reinhardtii
Method: EM (subtomogram averaging) / Resolution: 8.7 Å

EMDB-51802: In situ structure of peripheral stalk of mitochondrial ATP synthase from Chlamydomonas reinhardtii, consensus subtomogram average
Method: EM (subtomogram averaging) / Resolution: 5.1 Å

EMDB-51804: In situ structure of cytoplasmic microtubule of Chlamydomonas reinhardtii
Method: EM (subtomogram averaging) / Resolution: 4.7 Å

EMDB-51847: 80S Ribosome Average for EMPIAR-11830
Method: EM (subtomogram averaging) / Resolution: 4.0 Å

EMDB-51848: RuBisCo Average for EMPIAR-11830
Method: EM (subtomogram averaging) / Resolution: 7.5 Å

Source
  • Chlamydomonas reinhardtii (plant)

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