Extended Data Fig. 2: Cryo-EM structure determination of the CRL poly-ubiquitin chain formation complex.
From: Mechanism of millisecond Lys48-linked poly-ubiquitin chain formation by cullin-RING ligases
a, Flow chart describing the processing of the cryo-EM dataset (also see Table 1). The consensus map was refined to 3.76 Å. Additionally, five distinct focused maps were generated, with masking on the following subunits and the indicated residue ranges: (1) CUL2-Elongin B/C-FEM1C (residues 404-C)-RBX1 (residues 5-35); (2) FEM1C (residues N-404)-UBE2R2~UBD-UBA-Sil1 peptide-RBX1 (residues 35-104); (3) FEM1C (residues 150-404)-UBE2R2~UBD-UBA-Sil1-RBX1 (residues 35-104); (4) CUL2 (residues 429-556)-FEM1C (residues N-404)-UBE2R2~UBD-UBA-Sil1-RBX1 (residues 30-104); and (5) CUL2 (residues 429-556)-FEM1C (residues 150-404)-UBE2R2~UBD-UBA-Sil1-RBX1 (residues 30-104). N or C refer to N- or C-terminal residues. b, Final image of the composite cryo-EM map generated by merging all of the focused maps. c, Cryo-EM density of the consensus map as colored by the indicated local resolution. d, Graph plotting the Fourier shell correlation (FSC) versus inverse resolution (3.76 Å resolution at FSC = 0.143). e, Angular distribution of the consensus map. UBD, donor ubiquitin; UBA, acceptor ubiquitin.
