TABLE 2.
Composition of rHDL made with WT apoA-I or one of the mutant apoA-I and the conformational and stability characteristics of apoA-I in the rHDL
| POPC:Cholesterol:apoA-Ia (M:M:M) | α-Helixb (%) | T1/2c (°C) | ΔGDood (kcal/mol) | md [kcal/(moles of apoA-I × moles of GdnHCl)] | D1/2d (M) | |
| WT apoA-I | (76 ± 7):(7 ± 0.8):1 | 79 ± 2 | 78 ± 0.5 | 3.8 ± 0.3 | 1.4 ± 0.1 | 2.5 ± 0.1 |
| apoA-I[R123A] | (68 ± 5):(6 ± 0.7):1 | 77 ± 3 | 79 ± 1.0 | 3.4 ± 0.2 | 1.3 ± 0.1 | 2.6 ± 0.1 |
| apoA-I [R123E] | (71 ± 6):(7 ± 0.8):1 | 76 ± 1 | 79 ± 1.0 | 3.4 ± 0.2 | 1.4 ± 0.1 | 2.5 ± 0.0 |
| apoA-I [R131A] | (77 ± 4):(8 ± 0.5):1 | 79 ± 2 | 79 ± 0.5 | 3.9 ± 0.2 | 1.6 ± 0.1 | 2.4 ± 0.1 |
The values are mean ± SD from at least three experiments.
a
Molar composition of rHDL (determined in three independent reconstitution experiments).
b
Estimated from the [Θ222] at 25°C.
c
The midpoint of thermal unfolding, T1/2, at which half of the total change in the CD signal is observed, was determined from the thermal unfolding curves for rHDL.
d
The conformational stability, ΔGDo, m value, and midpoint of denaturation, D1/2, were determined by the linear extrapolation method from the chemical unfolding curves for rHDL.