Folding of cytochrome c initiated by submillisecond mixing.
Cytochrome c folding was initiated using a new solution mixer that provides a time window which covers over 90% of the burst phase unresolved by conventional stop-flow measurements. Folding was followed by resonance Raman scattering. Kinetic analysis of the high frequency Raman data indicates that a nascent phase occurs within the mixing dead time of 100 microseconds. A significant fraction of the protein was found to be trapped in a misfolded bis-histidine form during the nascent phase at pH 4.5, thereby preventing the protein from folding rapidly and homogeneously. The nascent phase was followed by a haem-ligand exchange phase that populates the native histidine-methionine coordinated form through a thermodynamically controlled equilibrium.
Takahashi
S
S
Institute of Physical and Chemical Research (RIKEN), Wako, Saitama, Japan.
Yeh
S R
SR
Das
T K
TK
Chan
C K
CK
Gottfried
D S
DS
Rousseau
D L
DL
D000818:Animals; D003574:Cytochrome c Group / Q000737:chemistry*; D006736:Horses; D007700:Kinetics; D009206:Myocardium / Q000201:enzymology; D011487:Protein Conformation; D017510:Protein Folding; D013059:Spectrum Analysis, Raman; D013816:Thermodynamics
D003574:Cytochrome c Group