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. 2012 Sep 21;40(21):11086–11099. doi: 10.1093/nar/gks871

Figure 2.

Figure 2.

RuvBL1 and RuvBL2 assemble as a dodecameric complex in the absence of tags. (A) SDS–PAGE of purified untagged RuvBL1–RuvBL2 complex evaluated by Coomassie staining (left) and western blot against histidine tag (right). Addition of TEV (TEV+) removes the tag of the His-RuvBL1–RuvBL2 complex (TEV−). (B) SEC of His–RuvBL1–RuvBL2 (solid line, TEV−) compared with untagged RuvBL1–RuvBL2 (dash line, TEV+). Molecular weight standards corresponding to 670, 440, 158 and 75 kDa are indicated. (C) BN–PAGE of His–RuvBL1–RuvBL2 (TEV−) compared with untagged RuvBL1–RuvBL2 (TEV+). The different oligomeric species are indicated. Detection of the histidine tag in BN–PAGE was performed by western blot. (D) EM of untagged RuvBL1–RuvBL2. Particles were collected, classified and averaged. 2D reference-free averages for the different views of the complex are shown. Compact and stretched conformations were observed. Scale bar, 10 nm.