Measuring the Conformational Stability of a Protein by NMR

Hydrogen Bonding Markedly Reduces the pK of Buried Carboxyl Groups in Proteins

“…The carboxyl oxygen atoms are almost completely buried for Asp33 and Asp76, but only ∼ 85% buried for Asp79 (Table 4). The continuous lines are curves calculated as described, 44 using the thermodynamic parameters given in Table 1.…”

“…Both methods have been described in detail. 44,45 The analysis of urea denaturation curves has also been described. 45 …”

“…Thermal denaturation experiments were done and the curves analyzed as described 44 using either an AVIV model 62DS or 202SF circular dichroism (CD) spectrometer (Aviv Instruments, Lakewood, NJ). The data were analyzed using Microcal Orgin 6.0 (Microcal Software, Inc., Northhampton, MA).…”

The HPr Proteins from the Thermophile Bacillus stearothermophilus can form Domain-swapped Dimers

“…The solvent-denaturation experiments, with urea as the denaturant, were performed in a similar fashion as the thermal unfolding experiments using methods as described. 41 …”

“…The urea-denaturation experiments were performed at 25 8C in 10 mM sodium phosphate buffer at pH 7.0 using techniques described earlier 41. b Thermal denaturation experiments were performed in 10 mM sodium phosphate buffer at pH 7.0.…”