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. 2007 Jan 3;104(2):467–472. doi: 10.1073/pnas.0603408104

Fig. 3.

Fig. 3.

Visualization of p97-Ufd1-Npl4-ubiquitin complexes by negative stain EM. (A) Band pass filtered images of the p97-Ufd1-Npl4-ubiquitin-GST complexes showing top views of p97 with a significant elongated extra-density at the periphery. (B) Class average of 150 p97-Ufd1-Npl4-ubiquitin-GST particles band-pass filtered to 25 Å and shown with superimposed isodensity contours. (C) Same average with overlaid yellow to mark the putative Ufd1-Npl4 complex, green for the ubiquitin-GST [Inset shows crystal structure of a GST dimer (PDB ID code 1PKZ) filtered to 25-Å resolution and viewed in an orientation matching the EM average] and in blue the p97 top view with the p97 N domain bound to the Ufd1-Npl4 complex. (D) Average of 150 top views of p97-Ufd1-Npl4ΔZF-ubiquitin-GST with superimposed density contours showing a similar domain organization to the Ufd1-Npl4-ubiquitin-GST bound to p97, in particular the GST moiety (bracket). (E) Gallery of p97-Ufd1-Npl4-ubiquitin (3–7 moieties, K48-linked) complexes incubated with an excess of purified IgGs (arrows) directed against ubiquitin. (Scale bar: 5 nm.)