Fig. 5: Molecular assembly of intact OGDHc.

a A representative complex containing tightly adjacent E1o and E3. The orange dot marked particle is identified as E1o, and the green triangle marked particle is identified as E3. b Organization of S. scrofa E2o and E1o domains. c Western blot analysis for subunit interaction detection between each OGDHc component. The first row is for anti-E1o detection and anti-E2o-ICD detection, and the second row is for anti-E3 detection. All of the experiments were performed in triplicate. Source data are provided in the Source Data file. d Surface plasmon resonance was used to detect the interaction between E1o and E3. The real-time binding curves were obtained by injecting different concentrations (49nm-25μM) of the analytes over the ligands-immobilized SPR chip. The curves were analyzed to calculate the equilibrium dissociation constant (KD = 1.84 μM). e Assembly of a structural model of intact eukaryotic OGDHC. The architecture model contains 6 binding modes (i) E1o-E2o_linker binding state, (ii) E1o-E2o_ICD binding state, (iii) E1o-E2o_LD binding state, iv)E3-E2o_liker binding state, v) E3-E2o_LD binding state, (vi) E1o-E3 binding state. Structure models were built on the basis of our single-particle E2o core (blue) in the center with E1o (orange, PDB:7WGR) and E3 (green, PDB:6I4Q) as peripheral subunit, LD (brown, PDB:1LAB) as E2o component. The flexible linker between the LD and the ICD in E2o and the N terminal linker in E1o is presented as the blue and yellow curves, respectively.