Quenched-flow HDX NMR None protection rate (s-1) ~20±5 SQAEFDKAAEEVKHLKTKPADEEMLFIYSHYKQATVGDINTERPGMLDFKGKAKWDAWNELKGTSKEDAMKAYIDKVEELKKKYGI

The time dependence of hydrogen exchange protection in the refolding of ACBP has been followed in 20 mM sodium acetate, 0.54 M GuHCl at pH 5.3 and 278 K. A total of 17 samples were prepared for a set of refolding times ranging from 0 ms to 250 ms.

Quenched-flow HDX NMR None Burst phase amplitude > 0.30 SQAEFDKAAEEVKHLKTKPADEEMLFIYSHYKQATVGDINTERPGMLDFKGKAKWDAWNELKGTSKEDAMKAYIDKVEELKKKYGI

The time dependence of hydrogen exchange protection in the refolding of ACBP has been followed in 20 mM sodium acetate, 0.54 M GuHCl at pH 5.3 and 278 K. A total of 17 samples were prepared for a set of refolding times ranging from 0 ms to 250 ms.

Quenched-flow HDX NMR None 0.20 < Burst phase amplitude < 0.30 SQAEFDKAAEEVKHLKTKPADEEMLFIYSHYKQATVGDINTERPGMLDFKGKAKWDAWNELKGTSKEDAMKAYIDKVEELKKKYGI

The time dependence of hydrogen exchange protection in the refolding of ACBP has been followed in 20 mM sodium acetate, 0.54 M GuHCl at pH 5.3 and 278 K. A total of 17 samples were prepared for a set of refolding times ranging from 0 ms to 250 ms.

Quenched-flow HDX NMR None 0.10 < Burst phase amplitude < 0.20 SQAEFDKAAEEVKHLKTKPADEEMLFIYSHYKQATVGDINTERPGMLDFKGKAKWDAWNELKGTSKEDAMKAYIDKVEELKKKYGI

The time dependence of hydrogen exchange protection in the refolding of ACBP has been followed in 20 mM sodium acetate, 0.54 M GuHCl at pH 5.3 and 278 K. A total of 17 samples were prepared for a set of refolding times ranging from 0 ms to 250 ms.

Native exchange NMR None log(P) > 4 SQAEFDKAAEEVKHLKTKPADEEMLFIYSHYKQATVGDINTERPGMLDFKGKAKWDAWNELKGTSKEDAMKAYIDKVEELKKKYGI

Amide hydrogen exchange with solvent deuterium was performed in deuterium oxide at pH 6.65. A sample of 2 mM free ACBP in H2O was adjusted to the desired pH and lyophilized repeatedly. After a final lyophilization the protein was dissolved in 600 ml 99.99% D2O and transferred to a precooled NMR tube. The protein-ligand sample was dissolved in 600 ml of D2O and 0.04 M potassium phosphate (pH 6.65) to concentrations of 2 mM. Potassium phosphate was deuterated by repeated lyophilization from D2O, redissolved in 99.99% D2O and quickly transferred to the lyophilized protein-ligand complex. After transfer to a precooled NMR tube the sample was immediately placed in the spectrometer which in advance had been tuned and calibrated on a similar sample.

Native exchange NMR None 2 < log(P) < 4 SQAEFDKAAEEVKHLKTKPADEEMLFIYSHYKQATVGDINTERPGMLDFKGKAKWDAWNELKGTSKEDAMKAYIDKVEELKKKYGI

Amide hydrogen exchange with solvent deuterium was performed in deuterium oxide at pH 6.65. A sample of 2 mM free ACBP in H2O was adjusted to the desired pH and lyophilized repeatedly. After a final lyophilization the protein was dissolved in 600 ml 99.99% D2O and transferred to a precooled NMR tube. The protein-ligand sample was dissolved in 600 ml of D2O and 0.04 M potassium phosphate (pH 6.65) to concentrations of 2 mM. Potassium phosphate was deuterated by repeated lyophilization from D2O, redissolved in 99.99% D2O and quickly transferred to the lyophilized protein-ligand complex. After transfer to a precooled NMR tube the sample was immediately placed in the spectrometer which in advance had been tuned and calibrated on a similar sample.