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. Author manuscript; available in PMC: 2015 Apr 27.
Published in final edited form as: J Am Chem Soc. 2013 Aug 27;135(36):13456–13463. doi: 10.1021/ja4053398

Figure 2.

Figure 2

Oligomerization pattern and nanopore conductance of pre-oligomerised ClyA nanopores. (a) Oligomerisation of ClyA nanopores examined by a 4-20% BN-PAGE. Proteins (1 mg/ml) were pre-incubated with 0.5% DDM for 30 min at 37 °C before loading into the gel (40 μg/lane). Lane 1: Protein ladder, lane 2: ClyA-WT, lane 3: ClyA-SS, lane 4: ClyA-AS and lane 5: ClyA-CS(b) Unitary nanopore conductance distribution obtained from 100 nanopores reconstituted in planar lipid bilayers for ClyA-WT (top), ClyA-CS (middle) and ClyA-AS (bottom) nanopores after pre-oligomerization in 0.5% DDM. Top: Type I ClyA-WT (GO = 1.81±0.04 nS) and Type II ClyA-WT (GO=2.21±0.07 nS) represented 24% and 19% of the inserted nanopores, respectively. Middle: ClyA-CS open pore conductance showed two clearly visible peaks. Type I ClyA-CS (GO=1.79±0.05 nS) included 35% and Type II ClyA-CS (2.23±0.08 nS) 23% of the reconstituted nanopores. Bottom: ClyA-AS nanopore conductance was relatively uniform with 52% of the reconstituted nanopores corresponding to Type I ClyA-AS (GO=1.80±0.05 nS) and 16% to Type II ClyA-AS (GO=2.24 ±0.09 nS).Recordings were carried out at −35 mV in 15 mM Tris.HCl, pH 7.5, 150mM NaCl and the temperature was 28°C.