Department of Physiology and Biophysics, Boston University School of Medicine, Boston, Massachusetts 02118, United States.
Biochemistry. 2011 May 17;50(19):3919-27. doi: 10.1021/bi200090y. Epub 2011 Apr 20.
Plasma lipoproteins are assemblies of lipids and apolipoproteins that mediate lipid transport and metabolism. High-density lipoproteins (HDL) remove excess cell cholesterol and provide protection against atherosclerosis. Important aspects of metabolic HDL remodeling, including apolipoprotein dissociation and lipoprotein fusion, are mimicked in thermal denaturation. We report the first study of the protein-lipid complexes by pressure perturbation calorimetry (PPC) beyond 100 °C. In PPC, volume expansion coefficient α(v)(T) is measured during heating; in proteins, α(v)(T) is dominated by hydration. Calorimetric studies of reconstituted HDL and of human high-density, low-density, and very low-density lipoproteins reveal that apolipoprotein unfolding, dissociation, and lipoprotein fusion are endothermic transitions without detectable volume changes. This may result from the limited applicability of PPC to slow kinetically controlled transitions such as thermal remodeling of lipoproteins and/or from the possibility that this remodeling causes no significant changes in the solvent structure and, hence, may not involve large transient solvent exposure of apolar moieties. Another conclusion is that apolipoprotein A-I in solution adsorbs to the phospholipid surface; protein hydration is preserved upon such adsorption. We posit that adsorption to a phospholipid surface helps recruit free apolipoprotein to the plasma membrane and facilitate HDL biogenesis.
血浆脂蛋白是脂质和载脂蛋白的复合物,介导脂质运输和代谢。高密度脂蛋白(HDL)可清除多余的细胞胆固醇,提供对动脉粥样硬化的保护。热变性模拟了代谢性 HDL 重塑的重要方面,包括载脂蛋白的解离和脂蛋白的融合。我们报告了首次在 100°C 以上通过压力扰动量热法(PPC)研究蛋白质-脂质复合物的研究。在 PPC 中,在加热过程中测量体积膨胀系数α(v)(T);在蛋白质中,α(v)(T)主要由水合作用决定。对重组 HDL 和人高密度脂蛋白、低密度脂蛋白和极低密度脂蛋白的量热研究表明,载脂蛋白的展开、解离和脂蛋白融合是吸热转变,没有可检测到的体积变化。这可能是由于 PPC 对动力学控制的缓慢转变(如脂蛋白的热重塑)的适用性有限,或者可能是这种重塑不会导致溶剂结构发生显著变化,因此可能不涉及对非极性部分的瞬时溶剂暴露。另一个结论是,溶液中的载脂蛋白 A-I 吸附到磷脂表面;在这种吸附过程中,蛋白质的水合作用得以保留。我们假设,吸附到磷脂表面有助于将游离载脂蛋白募集到质膜,并促进 HDL 的生物发生。