Membrane protein sequestering by ionic protein-lipid interactions.
Membrane protein sequestering by ionic protein-lipid interactions.
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DOI：
10.1038/nature10545
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发表时间：
2011-10-23
期刊：
影响因子：
64.8
通讯作者：
Jahn, Reinhard
中科院分区：
文献类型：
--
作者：
van den Bogaart, Geert;Meyenberg, Karsten;Risselada, H. Jelger;Amin, Hayder;Willig, Katrin I.;Hubrich, Barbara E.;Dier, Markus;Hell, Stefan W.;Grubmueller, Helmut;Diederichsen, Ulf;Jahn, Reinhard
神经元胞吐由SNARE蛋白syntaxin-1A催化。突触融合蛋白-1A在突触囊泡进行胞吐作用的位点处聚集在质膜中。然而，syntaxin-1A是如何被隔离的尚不清楚。在这里，我们表明，突触融合蛋白集群介导的静电相互作用与强阴离子脂质磷脂酰肌醇-4，5-二磷酸（PIP 2）。我们用超分辨率STED显微镜在PC 12细胞的质膜上发现，PIP 2是约73 nm大小的微区中的主要小叶内脂质。这种PIP 2的高积累是突触融合蛋白-1A隔离所必需的，因为磷酸酶synaptojanin-1对PIP 2的破坏减少了突触融合蛋白-1A的聚集。此外，共同重建的PIP 2和突触融合蛋白-1A的C-末端部分在人工巨大的单层囊泡导致分离的PIP 2和突触融合蛋白-1A到不同的领域，即使胆固醇是不存在的。我们的研究结果表明，静电蛋白质-脂质相互作用可以导致独立于胆固醇或脂质相的微区的形成。
Neuronal exocytosis is catalyzed by the SNARE protein syntaxin-1A. Syntaxin-1A is clustered in the plasma membrane at sites where synaptic vesicles undergo exocytosis. However, how syntaxin-1A is sequestered is unknown. Here, we show that syntaxin clustering is mediated by electrostatic interactions with the strongly anionic lipid phosphatidylinositol-4,5-bisphosphate (PIP2). We found with super-resolution STED microscopy on the plasma membrane of PC12 cells that PIP2 is the dominant inner-leaflet lipid in ~73 nm-sized microdomains. This high accumulation of PIP2 was required for syntaxin-1A sequestering, as destruction of PIP2 by the phosphatase synaptojanin-1 reduced syntaxin-1A clustering. Furthermore, co-reconstitution of PIP2 and the C-terminal part of syntaxin-1A in artificial giant unilamellar vesicles resulted in segregation of PIP2 and syntaxin-1A into distinct domains even when cholesterol was absent. Our results demonstrate that electrostatic protein-lipid interactions can result in the formation of microdomains independent of cholesterol or lipid phases.
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