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. Author manuscript; available in PMC: 2009 Oct 7.
Published in final edited form as: Science. 2008 Aug 7;321(5894):1354–1357. doi: 10.1126/science.1161070

Figure 3.

Figure 3

CHMP2ΔC-CHMP3 tube disassembly by VPS4B.

(A) Negative staining EM of tubes formed by CHMP2AΔC and CHMP3.

(B) Sucrose gradient analysis of CHMP2AΔC; (C) CHMP3; (D) VPS4B; (E) CHMP2AΔC-CHMP3-VPS4B complex formation (F) CHMP2AΔC-CHMP3-VPS4B complexes after incubation with ATP Mg2+.

(G) Negative staining EM of CHMP2AΔC-CHMP3 tubes revealing VPS4B on the inside.

(H) Radial density profile of a CHMP2AΔC-CHMP3-VPS4B and (I) of CHMP2AΔC-CHMP3 tubes calculated across the cross section of the tube.

(J) Negative staining EM after adding ATP Mg2+ to CHMP2AΔC-CHMP3-VPS4B tubes.

(K) Disassembly of fluorescein-labeled CHMP2AΔC-CHMP3 tubes measured by change in emission intensity upon addition of (i) HBS (magenta), (ii) 10 μM VPS4B (blue), (iii) 10 μM VPS4B plus 100μM AMP-PNP Mg2+ (green), (iv) 5 μM VPS4B plus 50μM ATP Mg2+ (black) and, (v) 10 μM VPS4B plus 100μM ATP Mg2+ (red). Inset shows the fluorescein-labeled CHMP2AΔC and CHMP3 visualized on an SDS-PAGE. (Scale bars 100 nm).