Εξειδίκευση τύπου :	Άρθρο σε επιστημονικό περιοδικό
Τίτλος:	Cryo-EM of a heterogeneous biochemical fraction elucidates multiple protein complexes from a multicellular thermophilic eukaryote
Δημιουργός/Συγγραφέας:	Semchonok, Dmitry A Kyrilis, Fotis L Hamdi, Farzad [EL] Καστρίτης, Παναγιώτης[EN] Kastritis, Panagiotis
Εκδότης:	Elsevier
Ημερομηνία:	2023-08-09
Γλώσσα:	Αγγλικά
ISSN:	25901524
DOI:	10.1016/j.yjsbx.2023.100094
Άλλο:	37638207
Περίληψη:	Biomolecular complexes and their interactions govern cellular structure and function. Understanding their architecture is a prerequisite for dissecting the cell's inner workings, but their higher-order assembly is often transient and challenging for structural analysis. Here, we performed cryo-EM on a single, highly heterogeneous biochemical fraction derived from Chaetomium thermophilum cell extracts to visualize the biomolecular content of the multicellular eukaryote. After cryo-EM single-particle image processing, results showed that a simultaneous three-dimensional structural characterization of multiple chemically diverse biomacromolecules is feasible. Namely, the thermophilic, eukaryotic complexes of (a) ATP citrate-lyase, (b) Hsp90, (c) 20S proteasome, (d) Hsp60 and (e) UDP-glucose pyrophosphorylase were characterized. In total, all five complexes have been structurally dissected in a thermophilic eukaryote in a total imaged sample area of 190.64 μm2, and two, in particular, 20S proteasome and Hsp60, exhibit side-chain resolution features. The C. thermophilum Hsp60 near-atomic model was resolved at 3.46 Å (FSC = 0.143) and shows a hinge-like conformational change of its equatorial domain, highly similar to the one previously shown for its bacterial orthologue, GroEL. This work demonstrates that cryo-EM of cell extracts will greatly accelerate the structural analysis of cellular complexes and provide unprecedented opportunities to annotate architectures of biomolecules in a holistic approach.
Τίτλος πηγής δημοσίευσης:	Journal of structural biology: X
Τόμος/Κεφάλαιο:	8
Σελίδες:	10
Θεματική Κατηγορία:	[EL] Δομική Βιολογία[EN] Structural Biology [EL] Βιοχημεία[EN] Biochemistry [EL] Βιοπληροφορική[EN] Bioinformatics [EL] Εφαρμοσμένη οπτική. Φωτονική[EN] Applied optics. Photonics
Λέξεις-Κλειδιά:	Biochemical fraction Cryo-EM Native cell extracts Simultaneous 3D reconstructions Single-particle image processing
Ηλεκτρονική διεύθυνση στον εκδότη (link):	https://www.sciencedirect.com/science/article/pii/S2590152423000107?via%3Dihub
Εμφανίζεται στις συλλογές:	Ινστιτούτο Χημικής Βιολογίας - Επιστημονικό έργο