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-Structure paper
タイトル | Integrative structure of a 10-megadalton eukaryotic pyruvate dehydrogenase complex from native cell extracts. |
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ジャーナル・号・ページ | Cell Rep, Vol. 34, Issue 6, Page 108727, Year 2021 |
掲載日 | 2021年2月9日 |
著者 | Fotis L Kyrilis / Dmitry A Semchonok / Ioannis Skalidis / Christian Tüting / Farzad Hamdi / Francis J O'Reilly / Juri Rappsilber / Panagiotis L Kastritis / |
PubMed 要旨 | The pyruvate dehydrogenase complex (PDHc) is a giant enzymatic assembly involved in pyruvate oxidation. PDHc components have been characterized in isolation, but the complex's quaternary structure ...The pyruvate dehydrogenase complex (PDHc) is a giant enzymatic assembly involved in pyruvate oxidation. PDHc components have been characterized in isolation, but the complex's quaternary structure has remained elusive due to sheer size, heterogeneity, and plasticity. Here, we identify fully assembled Chaetomium thermophilum α-keto acid dehydrogenase complexes in native cell extracts and characterize their domain arrangements utilizing mass spectrometry, activity assays, crosslinking, electron microscopy (EM), and computational modeling. We report the cryo-EM structure of the PDHc core and observe unique features of the previously unknown native state. The asymmetric reconstruction of the 10-MDa PDHc resolves spatial proximity of its components, agrees with stoichiometric data (60 E2p:12 E3BP:∼20 E1p: ≤ 12 E3), and proposes a minimum reaction path among component enzymes. The PDHc shows the presence of a dynamic pyruvate oxidation compartment, organized by core and peripheral protein species. Our data provide a framework for further understanding PDHc and α-keto acid dehydrogenase complex structure and function. |
リンク | Cell Rep / PubMed:33567276 |
手法 | EM (単粒子) |
解像度 | 6.9 - 32.7 Å |
構造データ | EMDB-12181: Cryo-EM map of Icosahedrally averaged native core of Pyruvate Dehydrogenase Complex from Ch. thermophilum EMDB-12233: EMDB-12234: |
由来 |
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キーワード | TRANSFERASE / Dihydrolipoyl / transacetylase / E2 / Pyruvate |