Thermodynamical properties of reaction intermediates during apoplastocyanin folding in time domain.
Two intermediates observed for the folding process of apoplastocyanin (apoPC) were investigated by using a photoinduced triggering system combined with the transient grating and transient lens methods. The thermodynamic quantities, enthalpy, heat capacity, partial volume, and thermal expansion volume changes during the protein folding reaction were measured in time domain for the first time. An interesting observation is the positive enthalpy changes during the folding process. This positive enthalpy change must be compensated by positive entropy changes, which could be originated from the dehydration effect of hydrophobic residues and/or the translational entropy gain of bulk water molecules. Observed negative heat capacity change was explained by the dehydration effect of hydrophilic residues and/or motional confinement of amino acid side chains and water molecules in apoPC. The signs of the volume change and thermal expansion volume were different for two processes and these changes were interpreted in terms of the different relative contributions of the hydration and the dehydration of the hydrophilic residues. These results indicated two-step hydrophobic collapses in the early stage of the apoPC folding, but the nature of the dynamics was different.
Baden
N
N
Department of Chemistry, Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan.
Hirota
S
S
Takabe
T
T
Funasaki
N
N
Terazima
M
M
D000596:Amino Acids / Q000737:chemistry; D001059:Apoproteins / Q000737:chemistry*; D002627:Chemistry, Physical / Q000379:methods*; D004058:Diffusion; D006358:Hot Temperature; D008027:Light; D010970:Plastocyanin / Q000737:chemistry*; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry; D013696:Temperature; D013816:Thermodynamics
D000596:Amino Acids; D001059:Apoproteins; D011506:Proteins; C040778:apoplastocyanin; D010970:Plastocyanin