The C2 domains of otoferlin, dysferlin, and myoferlin alter the packing of lipid bilayers.
The C2 domains of otoferlin, dysferlin, and myoferlin alter the packing of lipid bilayers.
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DOI：
10.1021/bi400432f
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发表时间：
2013-08-20
期刊：
影响因子：
2.9
通讯作者：
Johnson, Colin P.
中科院分区：
文献类型：
--
作者：
Marty, Naomi J.;Holman, Chelsea L.;Abdullah, Nazish;Johnson, Colin P.
费林蛋白是参与膜融合和分裂事件的大的多C2结构域膜蛋白。在这项研究中，我们调查的otoferlin，dysferlin和myoferlin的C2结构域的结合后的脂质双层的结构的影响。荧光测量表明，多C2结构域的肌电蛋白，dysferlin和otoferlin的结构改变了小的单层囊泡和巨大的质膜囊泡的脂质包装。这些蛋白质的活性在钙的存在下增强，并且需要带负电荷的脂质如磷脂酰丝氨酸或磷脂酰甘油的活性。对单个结构域的实验揭示了与dysferlin和myoferlin相比，otoferlin的C2A结构域之间的功能差异，并且截短研究表明每个后续C2结构域对脂质排序的影响似乎是累加的。最后，我们证明，这些蛋白质的膜上的活动是不敏感的高盐浓度，这表明一个非静电组件之间的相互作用的ferlin C2结构域和脂质双层。总之，这些数据表明，dysferlin，otoferlin和myoferlin不仅被动吸附到膜上，而且主动塑造脂质双层，这将导致高度弯曲或扭曲的膜区域，这可能有助于膜融合，裂变或招募其他膜运输蛋白。
Ferlins are large multi-C2 domain membrane proteins involved in membrane fusion and fission events. In this study we investigate the effects binding of the C2 domains of otoferlin, dysferlin and myoferlin have upon the structure of lipid bilayers. Fluorescence measurements indicate that multi-C2 domain constructs of myoferlin, dysferlin and otoferlin change the lipid packing of both small unilamellar vesicles and giant plasma membrane vesicles. The activities of these proteins were enhanced in the presence of calcium, and required negatively charged lipids like phosphatidylserine or phosphatidylglycerol for activity. Experiments on individual domains uncovered functional differences between the C2A domain of otoferlin as compared to dysferlin and myoferlin, and truncation studies suggest that the effects of each subsequent C2 domain on lipid ordering appear additive. Finally, we demonstrate that the activities of these proteins on membranes are insensitive to high salt concentrations, suggesting a non-electrostatic component to the interaction between ferlin C2 domains and lipid bilayers. Together, the data indicate that dysferlin, otoferlin, and myoferlin do not merely passively adsorb to membranes, but actively sculpt lipid bilayers, which would result in highly curved or distorted membrane regions that could facilitate membrane fusion, fission, or recruitment of other membrane trafficking proteins.
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