Structural transitions of Cthermo. (A)
Mg2+-titration monitored by CD at 260 and 287 nm at 37°C.
The KMg and n values are the same at both
wavelengths. (B) Urea titrations at Mg2+
concentrations of 0.2 (black), 0.4 (red), 0.6 (green), and 0.8 mM
(blue). All experiments were performed in 20 mM Tris⋅HCl, pH 8.1,
0.3 μM RNA, 37°C. (Inset) Stability extrapolated to
0 M urea vs. RTln[Mg2+]. The slope of this
plot is the Hill constant, n = 7.8 ± 0.3.
(C) Temperature melting at the indicated, constant
Mg2+ concentration (in mM) monitored by CD at 287 nm.
(D) Temperature dependence of the Mg2+
midpoint for the observed I-to-N transition. KMg was
obtained either from Mg2+-titrations at fixed temperature
(circles) or from temperature melting at fixed Mg2+
concentrations (triangles). The heat capacity change,
ΔCp, was obtained from the temperature-dependent
stability, ΔG(T) = ΔH* − TΔS* + ΔCp[(T
− 300) − T ln(T/300)], where ΔH* and ΔS* are the enthalpy
and entropy changes at 300 K.