Figure 4.

Reverse-phase HPLC chromatograms of the sample solutions (with AEMTS blocking) obtained from the temperature-jump experiment for the folding intermediates of HEL generated by the reaction of R with three equivalents of DHS^ox at pH 8.0 in the presence of 2 M urea. Reaction conditions were [R]₀ = [DHS^ox]₀/3 = 9.8 μM. (a) Incubated at 5 °C for 300 min; (b) Incubated at 15 °C for 20 min after (a); (c) Incubated at 25 °C for 20 min after (b); (d) Incubated at 35 °C for 20 min after (c); (e) Incubated at 45 °C for 20 min after (d); (f) Incubated at 5 °C for 300 min after (e). See the text for details of the HPLC analysis conditions.