Figure 1. ΔN-VAT unfolds other copies of ΔN-VAT.

(a) Cartoon showing VAT (purple ring) working in concert with the 20S proteasome (blue and pink rings) to degrade protein substrates (yellow). (b) An SDS-PAGE gel showing that both ΔN-VAT and pyruvate kinase (part of the ATP regenerating system) are degraded in an ATP and 20S proteasome dependent manner. (c) Quantification of the self-unfolding reaction. All measurements done in triplicate.(d) Representative section of a micrograph of ΔN-VAT in the presence of 5 mM ADP, corresponding to ~25% of the full field-of-view. Inset shows the power spectrum from a fibril, with a characteristic layer line at ~1/55 Å⁻¹. (e) Representative section of a micrograph of ΔN-VAT in the presence of 5 mM ATPγS. Example single particles are circled in red. Scale bar, 500 Å. (f) 2D class average images showing views of single particles of ΔN-VAT in the presence of ATPγS.

DOI: http://dx.doi.org/10.7554/eLife.25754.003

Figure 1—figure supplement 1. Domain arrangement and oligomeric structure of VAT.

Cartoon representation of the VAT primary structure (a) protomer structure (b) and hexamer structure (c). In (c) the stacked ring structure is shown with the AAA+ nucleotide binding domains colored in blue and green for NBD1 and NBD2, respectively, with the N-terminal domain (NTD: orange) co-planar with the NBD1 rings.

DOI: http://dx.doi.org/10.7554/eLife.25754.004