Pulse labeling HDX NMR None protection within 3.5 ms KVFERCELARTLKRLGMDGYRGISLANWMCLAKWESGYNTRATNYNAGDRSTDYGIFQINSRYWCNDGKTPGAVNACHLSCSALLQDNIADAVACAKRVVRDPQGIRAWVAWRNRCQNRDVRQYVQGCGV

A QFM5 module was used to dilute a solution of human lysozyme in 6 M GuDCl /D2O (1 vol) with 20 mM sodium acetate in H2O, pH 5.5 (10 vol), to initiate the refolding reaction. The intrinsic rate of amide hydrogen exchange at 20 °C at the final refolding pH of 5.3 is approximately 10 s, and thus no significant exchange of amide deuterons occurs over the time course of folding (0-500 ms) monitored in this phase. At different times following the initiation of refolding, the pH of the refolding medium was jumped to 9.3 by dilution with 200 mM sodium borate buffer at pH 10.0 (5 volumes); this labeling pulse was applied for 8 ms.

Pulse labeling HDX NMR None protection within 70 ms KVFERCELARTLKRLGMDGYRGISLANWMCLAKWESGYNTRATNYNAGDRSTDYGIFQINSRYWCNDGKTPGAVNACHLSCSALLQDNIADAVACAKRVVRDPQGIRAWVAWRNRCQNRDVRQYVQGCGV

A QFM5 module was used to dilute a solution of human lysozyme in 6 M GuDCl /D2O (1 vol) with 20 mM sodium acetate in H2O, pH 5.5 (10 vol), to initiate the refolding reaction. The intrinsic rate of amide hydrogen exchange at 20 °C at the final refolding pH of 5.3 is approximately 10 s, and thus no significant exchange of amide deuterons occurs over the time course of folding (0-500 ms) monitored in this phase. At different times following the initiation of refolding, the pH of the refolding medium was jumped to 9.3 by dilution with 200 mM sodium borate buffer at pH 10.0 (5 volumes); this labeling pulse was applied for 8 ms.

Pulse labeling HDX NMR None protection around 100 ms KVFERCELARTLKRLGMDGYRGISLANWMCLAKWESGYNTRATNYNAGDRSTDYGIFQINSRYWCNDGKTPGAVNACHLSCSALLQDNIADAVACAKRVVRDPQGIRAWVAWRNRCQNRDVRQYVQGCGV

A QFM5 module was used to dilute a solution of human lysozyme in 6 M GuDCl /D2O (1 vol) with 20 mM sodium acetate in H2O, pH 5.5 (10 vol), to initiate the refolding reaction. The intrinsic rate of amide hydrogen exchange at 20 °C at the final refolding pH of 5.3 is approximately 10 s, and thus no significant exchange of amide deuterons occurs over the time course of folding (0-500 ms) monitored in this phase. At different times following the initiation of refolding, the pH of the refolding medium was jumped to 9.3 by dilution with 200 mM sodium borate buffer at pH 10.0 (5 volumes); this labeling pulse was applied for 8 ms.