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. Author manuscript; available in PMC: 2013 Sep 13.
Published in final edited form as: J Mol Biol. 2012 Mar 3;419(0):89–101. doi: 10.1016/j.jmb.2012.02.043

Figure 5.

Figure 5

Dissociation temperature during SDS-PAGE separation for a) wild type, b) mutant G57A/G59A, c) mutant G57S/G57S, and d) mutant R100L/G135W. Wild type and mutant OmpF proteins were expressed, purified, and refolded from inclusion bodies as described in Methods. Folding reactions were quenched by adding 5× SDS gel-loading buffer [45] to a final dilution of 1× SDS gel loading buffer. 50 μl of sample was loaded on a 4-20% acrylamide continous gradient precast gel to resolve the folded and unfolded populations [12]. Wild type and mutant proteins were subjected to temperature increases of 2°C from 20 to 90°C. Gel lanes at the transition temperature are in boxes. The dissociation of dimeric species takes place first followed by unfolding of the monomeric species. Trimeric (T), Dimeric (D), Monomeric (M), and unfolded (U) species are labeled.