鉴定溶酶体中尼曼-匹克 C2 向 NPC1 转移胆固醇所必需的表面残基。
Identification of surface residues on Niemann-Pick C2 essential for hydrophobic handoff of cholesterol to NPC1 in lysosomes.
机构信息
Department of Molecular Genetics, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75390, USA.
出版信息
Cell Metab. 2010 Aug 4;12(2):166-73. doi: 10.1016/j.cmet.2010.05.016.
Abstract
Water-soluble Niemann-Pick C2 (NPC2) and membrane-bound NPC1 are cholesterol-binding lysosomal proteins required for export of lipoprotein-derived cholesterol from lysosomes. The binding site in NPC1 is located in its N-terminal domain (NTD), which projects into the lysosomal lumen. Here we perform alanine-scanning mutagenesis to identify residues in NPC2 that are essential for transfer of cholesterol to NPC1(NTD). Transfer requires three residues that form a patch on the surface of NPC2. We previously identified a patch of residues on the surface of NPC1(NTD) that are required for transfer. We present a model in which these two surface patches on NPC2 and NPC1(NTD) interact, thereby opening an entry pore on NPC1(NTD) and allowing cholesterol to transfer without passing through the water phase. We refer to this transfer as a hydrophobic handoff and hypothesize that this handoff is essential for cholesterol export from lysosomes.
摘要
水溶性神经鞘氨醇胆固醇酰基转移酶 2（NPC2）和膜结合 NPC1 是胆固醇结合溶酶体蛋白，对于从溶酶体中输出脂蛋白衍生的胆固醇是必需的。NPC1 中的结合位点位于其 N 端结构域（NTD），该结构域向溶酶体腔中突出。在这里，我们进行丙氨酸扫描诱变，以鉴定 NPC2 中对 NPC1（NTD）胆固醇转移至关重要的残基。转移需要形成 NPC2 表面斑块的三个残基。我们之前鉴定了 NPC1（NTD）表面上的一个残基斑块，该残基斑块对于转移是必需的。我们提出了一个模型，其中 NPC2 和 NPC1（NTD）的这两个表面斑块相互作用，从而在 NPC1（NTD）上打开一个入口孔，允许胆固醇转移而无需穿过水相。我们将这种转移称为疏水传递，并假设这种传递对于胆固醇从溶酶体中输出是必需的。
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