The cold denatured state is compact but expands at low temperatures: hydrodynamic properties of the cold denatured state of the C-terminal domain of L9.
A point mutation of a small globular protein, the C-terminal domain of L9 destabilizes the protein and leads to observable cold-denaturation at temperatures above zero. The cold denatured state is in slow exchange with the native state on the NMR time scale, and this allows the hydrodynamic properties of the cold unfolded state and the native state to be measured under identical conditions using pulsed-field gradient NMR diffusion measurements. This provides the first experimental measurement of the hydrodynamic properties of a cold unfolded protein and its folded form under identical conditions. Hydrodynamic radii of the cold-induced unfolded states were measured for a set of temperatures ranging from 2 degrees C to 25 degrees C at pD 6.6 in the absence of denaturant. The cold unfolded state is compact compared to the urea or acid unfolded state and a trend of increasing radii of hydration is observed as the temperature is lowered. These observations are confirmed by experiments on the same protein at pD 8.0, where it is more stable, in the presence of a modest concentration of urea. The expansion of the cold-denatured state at lower temperatures is consistent with the temperature dependence of hydrophobic interactions.
Li
Ying
Y
Department of Chemistry, State University of New York at Stony Brook, Stony Brook, NY 11794-3400, USA.
Shan
Bing
B
Raleigh
Daniel P
DP
D000409:Alanine / Q000235:genetics / Q000502:physiology; D019943:Amino Acid Substitution; D002942:Circular Dichroism; D003080:Cold Temperature; D007532:Isoleucine / Q000235:genetics / Q000502:physiology; D008958:Models, Molecular; D050505:Mutant Proteins / Q000737:chemistry; D019906:Nuclear Magnetic Resonance, Biomolecular; D011489:Protein Denaturation; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D012269:Ribosomal Proteins / Q000737:chemistry* / Q000235:genetics; D013816:Thermodynamics
D050505:Mutant Proteins; D012269:Ribosomal Proteins; C049650:ribosomal protein L9; D007532:Isoleucine; D000409:Alanine