Figure 4 | Scientific Reports

Figure 4

From: Plasticity in the Oxidative Folding Pathway of the High Affinity Nerita Versicolor Carboxypeptidase Inhibitor (NvCI)

Figure 4

Reductive unfolding of native NvCI. Native NvCI was treated with: (a) 1 mM DTT in Tris-HCl (pH 8.4) and (b) 20 mM TCEP in sodium acetate (pH 4.5). Time course intermediates were trapped by acidification and analyzed by RP-HPLC. N and R stand for native and reduced NvCI; II for an intermediate that accumulates upon reduction with 20 mM TCEP. (c) Reductive unfolding of native NvCI followed by changes in tryptophan fluorescence spectra, after the addition of reducing agents. Native NvCI was reduced with TCEP (20 mM) in sodium acetate pH 4.5 (graph below) and DTT (1.0 and 20 mM) in Tris.Cl pH 8, 4 (graph above). Tryptophan fluorescence spectra were measured over time and the spectrum area was plotted as a function of time.

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