Gorshkova Irina N, Mei Xiaohu, Atkinson David
Department of Physiology and Biophysics, Boston University School of Medicine, Boston, MA 02118 Department of Medicine, Boston University School of Medicine, Boston, MA 02118.
Department of Physiology and Biophysics, Boston University School of Medicine, Boston, MA 02118.
J Lipid Res. 2014 Sep;55(9):1876-85. doi: 10.1194/jlr.M047241. Epub 2014 Jun 11.
We found earlier that apoA-I variants that induced hypertriglyceridemia (HTG) in mice had increased affinity to TG-rich lipoproteins and thereby impaired their catabolism. Here, we tested whether a naturally occurring human apoA-I mutation, Lys107del, associated with HTG also promotes apoA-I binding to TG-rich particles. We expressed apoA-I[Lys107del] variant in Escherichia coli, studied its binding to TG-rich emulsion particles, and performed a physicochemical characterization of the protein. Compared with WT apoA-I, apoA-I[Lys107del] showed enhanced binding to TG-rich particles, lower stability, and greater exposure of hydrophobic surfaces. The crystal structure of truncated, Δ(185-243), apoA-I suggests that deletion of Lys107 disrupts helix registration and disturbs a stabilizing salt bridge network in the N-terminal helical bundle. To elucidate the structural changes responsible for the altered function of apoA-I[Lys107del], we studied another mutant, apoA-I [Lys107Ala]. Our findings suggest that the registry shift and ensuing disruption of the inter-helical salt bridges in apoA-I[Lys107del] result in destabilization of the helical bundle structure and greater exposure of hydrophobic surfaces. We conclude that the structural changes in the apoA-I[Lys107del] variant facilitate its binding to TG-rich lipoproteins and thus, may reduce their lipolysis and contribute to the development of HTG in carriers of the mutation.
我们先前发现,在小鼠中诱导高甘油三酯血症(HTG)的载脂蛋白A-I(apoA-I)变体对富含甘油三酯的脂蛋白的亲和力增加,从而损害其分解代谢。在此,我们测试了一种与HTG相关的天然存在的人类apoA-I突变Lys107del是否也促进apoA-I与富含甘油三酯的颗粒结合。我们在大肠杆菌中表达了apoA-I[Lys107del]变体,研究了其与富含甘油三酯的乳剂颗粒的结合,并对该蛋白进行了物理化学表征。与野生型apoA-I相比,apoA-I[Lys107del]对富含甘油三酯的颗粒表现出更强的结合力、更低的稳定性以及更大程度的疏水表面暴露。截短的Δ(185 - 243) apoA-I的晶体结构表明,Lys107的缺失破坏了螺旋排列,并扰乱了N端螺旋束中的稳定盐桥网络。为了阐明导致apoA-I[Lys107del]功能改变的结构变化,我们研究了另一个突变体apoA-I [Lys107Ala]。我们的研究结果表明,apoA-I[Lys107del]中螺旋排列的改变以及随之而来的螺旋间盐桥的破坏导致螺旋束结构不稳定和疏水表面更大程度的暴露。我们得出结论,apoA-I[Lys107del]变体中的结构变化促进了其与富含甘油三酯的脂蛋白的结合,因此可能会减少它们的脂解作用,并导致该突变携带者发生HTG。