HDX-folding competition by NMR None strong protection within 3.5 ms KVFSKCELAHKLKAQEMDGFGGYSLANWVCMAEYESNFNTRAFNGKNANGSSDYGLFQLNNKWWCKDNKRSSSNACNIMCSKLLDENIDDDISCAKRVVRDPKGMSAWKAWVKHCKDKDLSEYLASCNL

Equine lysozyme denatured in 6 M GdnHCl was diluted rapidly with deuterated buffer solution at pH 7.5 and 25 °C, in order to initiate simultaneous hydrogen exchange and protein refolding. The intrinsic time constants for amide hydrogen exchange under these conditions are of the order of 10-200 ms. Exchange was allowed to occur for one minute, and was then quenched by decreasing the pH to 4.5.

HDX-folding competition by NMR None strong protection within 10 ms KVFSKCELAHKLKAQEMDGFGGYSLANWVCMAEYESNFNTRAFNGKNANGSSDYGLFQLNNKWWCKDNKRSSSNACNIMCSKLLDENIDDDISCAKRVVRDPKGMSAWKAWVKHCKDKDLSEYLASCNL

Equine lysozyme denatured in 6 M GdnHCl was diluted rapidly with deuterated buffer solution at pH 7.5 and 25 °C, in order to initiate simultaneous hydrogen exchange and protein refolding. The intrinsic time constants for amide hydrogen exchange under these conditions are of the order of 10-200 ms. Exchange was allowed to occur for one minute, and was then quenched by decreasing the pH to 4.5.

HDX-folding competition by NMR None strong protection within 50 ms KVFSKCELAHKLKAQEMDGFGGYSLANWVCMAEYESNFNTRAFNGKNANGSSDYGLFQLNNKWWCKDNKRSSSNACNIMCSKLLDENIDDDISCAKRVVRDPKGMSAWKAWVKHCKDKDLSEYLASCNL

Equine lysozyme denatured in 6 M GdnHCl was diluted rapidly with deuterated buffer solution at pH 7.5 and 25 °C, in order to initiate simultaneous hydrogen exchange and protein refolding. The intrinsic time constants for amide hydrogen exchange under these conditions are of the order of 10-200 ms. Exchange was allowed to occur for one minute, and was then quenched by decreasing the pH to 4.5.

Native exchange NMR None log(P) > 5 KVFSKCELAHKLKAQEMDGFGGYSLANWVCMAEYESNFNTRAFNGKNANGSSDYGLFQLNNKWWCKDNKRSSSNACNIMCSKLLDENIDDDISCAKRVVRDPKGMSAWKAWVKHCKDKDLSEYLASCNL

Samples for hydrogen exchange studies of the native holo state of equine lysozyme were adjusted to pH 4.5 in H2O and freeze-dried. The protein was then dissolved in D2O in the presence of 10 mM CaCl2, and the apparent pH adjusted to 4.5 using 2HCl and NaO2H. Samples were allowed to undergo exchange at 25 °C for time intervals ranging from a few minutes to 63 days. At the end of each exchange period, phase-sensitive COSY spectra were acquired.

Native exchange NMR None 4 < log(P) < 5 KVFSKCELAHKLKAQEMDGFGGYSLANWVCMAEYESNFNTRAFNGKNANGSSDYGLFQLNNKWWCKDNKRSSSNACNIMCSKLLDENIDDDISCAKRVVRDPKGMSAWKAWVKHCKDKDLSEYLASCNL

Samples for hydrogen exchange studies of the native holo state of equine lysozyme were adjusted to pH 4.5 in H2O and freeze-dried. The protein was then dissolved in D2O in the presence of 10 mM CaCl2, and the apparent pH adjusted to 4.5 using 2HCl and NaO2H. Samples were allowed to undergo exchange at 25 °C for time intervals ranging from a few minutes to 63 days. At the end of each exchange period, phase-sensitive COSY spectra were acquired.