Tail-Anchored Protein Insertion by a Single Get1/2 Heterodimer.

Tail-Anchored Protein Insertion by a Single Get1/2 Heterodimer.
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通过单个 Get1/2 异二聚体进行尾锚定蛋白插入。

DOI:
10.1016/j.celrep.2017.08.035
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发表时间:
2017
期刊:
影响因子:
8.8
通讯作者:
Keenan,RobertJ
Keenan,RobertJ
中科院分区:
生物学1区
文献类型:
--
作者:
Zalisko,BenjaminE;Chan,Charlene;Denic,Vladimir;Rock,RonaldS;Keenan,RobertJ

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Get 1/2跨膜复合物驱动尾锚定(TA)蛋白从胞质伴侣Get 3插入内质网膜。对Get 1/2如何协调这一过程的机械洞察力由于缺乏对复杂的基本架构的理解而受到混淆。在这里,我们定义的寡聚状态的全长Get 1/2在重构的脂质双层相结合的单分子和散装荧光测量定量体外插入分析。我们发现,一个单一的Get 1/2异二聚体是足够的插入,并证明保守的胞质区域的Get 1和Get 2结合不对称的Get 3同源二聚体的相对亚基。总之,我们的研究结果定义了一个简化的模型如何Get 1/2和Get 3协调TA蛋白插入。
The Get1/2 transmembrane complex drives the insertion of tail-anchored (TA) proteins from the cytosolic chaperone Get3 into the endoplasmic reticulum membrane. Mechanistic insight into how Get1/2 coordinates this process is confounded by a lack of understanding of the basic architecture of the complex. Here, we define the oligomeric state of full-length Get1/2 in reconstituted lipid bilayers by combining single-molecule and bulk fluorescence measurements with quantitative in vitro insertion analysis. We show that a single Get1/2 heterodimer is sufficient for insertion and demonstrate that the conserved cytosolic regions of Get1 and Get2 bind asymmetrically to opposing subunits of the Get3 homodimer. Altogether, our results define a simplified model for how Get1/2 and Get3 coordinate TA protein insertion.
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