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. 2010 Oct 1;21(19):3396–3408. doi: 10.1091/mbc.E10-06-0512

Figure 4.

Figure 4.

Vps4 oligomer MIT domain composition for ESCRT-III disassembly. (A) ESCRT-III disassembly reactions were performed with vps4Δ membranes (0.5 OD600 equivalents) and 100 nM Vps4, 100 nM Vps4ΔMIT, or 100 nM Vps4 with increasing concentrations of Vps4ΔMIT (100 nM, 500 nM, and 1 μM). (B) ESCRT-III release reactions were performed as in A but with increasing concentrations of Vps4ΔMIT,E233Q (100 nM, 500 nM, and 1 μM). (C) ATPase activity of 250 nM Vps4 alone or with addition of 1 μM or 2 μM Vps4ΔMIT,E233Q. The initial ATP concentration was 1 mM. Activity is presented as ADP generated per wild-type Vps4 per minute. (D) ESCRT-III disassembly reactions were performed as in A but with 30 nM Vps4 alone, 1.2 μM Vps4ΔMIT,E233Q alone, 30 nM Vps4 and 1.2 μM Vps4ΔMIT,E233Q, or 100 nM Vps4. The addition of Vps4ΔMIT,E233Q enhanced 30 nM Vps4-mediated ESCRT-III disassembly (p < 0.0002).