TY - JOUR
T1 - Structural analysis of an endogenous 4-megadalton succinyl-CoA-generating metabolon
AU - Skalidis, Ioannis
AU - Kyrilis, Fotis L.
AU - Tüting, Christian
AU - Hamdi, Farzad
AU - Träger, Toni K.
AU - Belapure, Jaydeep
AU - Hause, Gerd
AU - Fratini, Marta
AU - O’Reilly, Francis J.
AU - Heilmann, Ingo
AU - Rappsilber, Juri
AU - Kastritis, Panagiotis L.
N1 - Funding Information:
We would like to thank all the Kastritis lab members for fruitful discussions. This work was supported by the European Union through funding of the Horizon Europe ERA Chair “hot4cryo” project number 101086665 (to P.L.K.), the Federal Ministry of Education and Research (BMBF, ZIK program) (Grant nos. 03Z22HN23, 03Z22HI2 and 03COV04 to P.L.K.), the European Regional Development Funds (EFRE) for Saxony-Anhalt (Grant no. ZS/2016/04/78115 to P.L.K.), the Deutsche Forschungsgemeinschaft (project number 391498659, RTG 2467), and the Martin-Luther University of Halle-Wittenberg.
Publisher Copyright:
© 2023, The Author(s).
PY - 2023/5/22
Y1 - 2023/5/22
N2 - The oxoglutarate dehydrogenase complex (OGDHc) participates in the tricarboxylic acid cycle and, in a multi-step reaction, decarboxylates α-ketoglutarate, transfers succinyl to CoA, and reduces NAD+. Due to its pivotal role in metabolism, OGDHc enzymatic components have been studied in isolation; however, their interactions within the endogenous OGDHc remain elusive. Here, we discern the organization of a thermophilic, eukaryotic, native OGDHc in its active state. By combining biochemical, biophysical, and bioinformatic methods, we resolve its composition, 3D architecture, and molecular function at 3.35 Å resolution. We further report the high-resolution cryo-EM structure of the OGDHc core (E2o), which displays various structural adaptations. These include hydrogen bonding patterns confining interactions of OGDHc participating enzymes (E1o-E2o-E3), electrostatic tunneling that drives inter-subunit communication, and the presence of a flexible subunit (E3BPo), connecting E2o and E3. This multi-scale analysis of a succinyl-CoA-producing native cell extract provides a blueprint for structure-function studies of complex mixtures of medical and biotechnological value.
AB - The oxoglutarate dehydrogenase complex (OGDHc) participates in the tricarboxylic acid cycle and, in a multi-step reaction, decarboxylates α-ketoglutarate, transfers succinyl to CoA, and reduces NAD+. Due to its pivotal role in metabolism, OGDHc enzymatic components have been studied in isolation; however, their interactions within the endogenous OGDHc remain elusive. Here, we discern the organization of a thermophilic, eukaryotic, native OGDHc in its active state. By combining biochemical, biophysical, and bioinformatic methods, we resolve its composition, 3D architecture, and molecular function at 3.35 Å resolution. We further report the high-resolution cryo-EM structure of the OGDHc core (E2o), which displays various structural adaptations. These include hydrogen bonding patterns confining interactions of OGDHc participating enzymes (E1o-E2o-E3), electrostatic tunneling that drives inter-subunit communication, and the presence of a flexible subunit (E3BPo), connecting E2o and E3. This multi-scale analysis of a succinyl-CoA-producing native cell extract provides a blueprint for structure-function studies of complex mixtures of medical and biotechnological value.
UR - https://doi.org/10.1038/s42003-023-04885-0
U2 - 10.1038/s42003-023-04885-0
DO - 10.1038/s42003-023-04885-0
M3 - Article
C2 - 37217784
AN - SCOPUS:85159818685
SN - 2399-3642
VL - 6
JO - Communications Biology
JF - Communications Biology
IS - 1
M1 - 552
ER -