Oda Michael N, Forte Trudy M, Ryan Robert O, Voss John C
Children's Hospital Oakland Research Institute, Oakland, California 94609-1673, USA.
Nat Struct Biol. 2003 Jun;10(6):455-60. doi: 10.1038/nsb931.
Exchangeable apolipoproteins can convert between lipid-free and lipid-associated states. The C-terminal domain of human apolipoprotein A-I (apoA-I) plays a role in both lipid binding and self-association. Site-directed spin-label electron paramagnetic resonance spectroscopy was used to examine the structure of the apoA-I C terminus in lipid-free and lipid-associated states. Nitroxide spin-labels positioned at defined locations throughout the C terminus were used to define discrete secondary structural elements. Magnetic interactions between probes localized at positions 163, 217 and 226 in singly and doubly labeled apoA-I gave inter- and intramolecular distance information, providing a basis for mapping apoA-I tertiary and quaternary structure. Spectra of apoA-I in reconstituted HDL revealed a lipid-induced transition of defined random coils and beta-strands into alpha-helices. This conformational switch is analogous to triggered events in viral fusion proteins and may serve as a means to overcome the energy barriers of lipid sequestration, a critical step in cholesterol efflux and HDL assembly.
可交换载脂蛋白能够在无脂状态和与脂质结合的状态之间转换。人载脂蛋白A-I(apoA-I)的C末端结构域在脂质结合和自身缔合过程中均发挥作用。运用定点自旋标记电子顺磁共振波谱法研究了apoA-I C末端在无脂状态和与脂质结合状态下的结构。位于整个C末端特定位置的氮氧化物自旋标记用于确定离散的二级结构元件。在单标记和双标记的apoA-I中,定位在163、217和226位的探针之间的磁相互作用给出了分子间和分子内距离信息,为绘制apoA-I的三级和四级结构提供了依据。重组高密度脂蛋白(HDL)中apoA-I的光谱显示,脂质诱导特定的无规卷曲和β链转变为α螺旋。这种构象转换类似于病毒融合蛋白中的触发事件,可能是克服脂质隔离能量障碍的一种方式,脂质隔离是胆固醇外流和HDL组装中的关键步骤。