FigureS1.
Far-UV Circular Dichroic spectra of HubWA, I13V, and I36V. Solid line, dot line, and short-dash line represent HubWA, I13V,
and I36V, respectively. About 30 mM of protein sample was placed in a cell with 1 mm pathlength. The spectral measurements
were done at 25℃ by using a JASCO J-810 spectropolarimeter (Tokyo, Japan). The unit for molar residue ellipticity, [q], is
deg cm2 dmol-1 residue-1.
FigureS2.
Folding kinetics results of I13V and I36V. Panels A and C represent the rate profile of I13V and I36V, respectively. The rate
constants of the first, second and third phases in refolding measurements are shown in circles, squares, and triangles, respectively.
Closed diamonds in panel A and C represent the rate constant obtained from unfolding measurements. Panels B and D illustrate
the amplitudes of folding and unfolding measurements of I13V and I36V, respectively. Circles, squares and triangles represent
the amplitudes of the first, second and third refolding phases, respectively. Baseline values were added to each amplitude
value to show the trend of amplitude change in the same plot. Open diamonds represent the fluorescence signal at long time,
which are equivalent to the fluorescence signal in equilibrium folding transition. Closed diamonds in panel B and D are the
values obtained by addition of amplitudes of unfolding reaction and baseline values to show the equilibrium unfolding transition
and amplitude change in the same plot. The dotted lines in panels B and C are the base lines for native state (lower dotted
line) and unfolded state (upper dotted line). AU represents auxiliary unit.
FigureS3.
Far-UV Circular Dichroic spectra of HubWA and hydrophobic residue to valine variant proteins. Solid line, long-dash line,
medium-dash line, short-dash line, dot line, dash-dot line, dash-dot-dot line, medium-medium line, short-short-short line,
short-short line, and short-long line represent HubWA, I3V, L15V, I23V, I30V, L43V, L50V, L56V, I61V, L67V, and L69V, respectively.
About 30 mM of protein sample was placed in a cell with1 mm pathlength. The spectral measurements were done at 25℃ by using
a JASCO J-810 spectropolarimeter (Tokyo, Japan). The unit for molar residue ellipticity, [q], is deg cm2 dmol-1 residue-1.
