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. 2008 Sep;132(3):351–360. doi: 10.1085/jgp.200809984

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© 2008 Anderson and Blaustein

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Figure 8. — Intersubunit dimer formation in S312C channels. Lanes 1 and 2: At pH 8.5 under denaturing conditions, the prepore form of (PA₆₃)₇ dissociates into a single monomeric band on SDS-PAGE (lane 1, S312C; lane 2, S330C). Samples in lanes 1 and 2 were not exposed to liposomes. Lane 3: At pH 6.5, under oxidizing conditions (0.3% H₂O₂), heptameric S312C (PA₆₃)₇ prepore converts to the pore form and resolves as an SDS-resistant complex on SDS-PAGE. Lane 4: Upon boiling in 8 M urea, the S312C heptamer dissociates to monomeric and dimeric species. Lane 5: If the sample is boiled in the presence of 10 mM DTT, only monomeric S312C PA₆₃ is seen. Lanes 6–8: Similar analysis of S330C, predicted to point away from the pore, yields no evidence of intersubunit dimer formation.