Hariri Hanaa, Bhattacharya Nilakshee, Johnson Kerri, Noble Alex J, Stagg Scott M
Institute of Molecular Biophysics, Florida State University, Tallahassee, FL 32306, USA.
Department of Chemistry and Biochemistry, Florida State University, Tallahassee, FL 32306, USA.
J Mol Biol. 2014 Nov 11;426(22):3811-3826. doi: 10.1016/j.jmb.2014.08.023. Epub 2014 Sep 2.
The small GTPase protein Sar1 is known to be involved in both the initiation of COPII-coated vesicle formation and scission of the nascent vesicle from the endoplasmic reticulum. The molecular details for the mechanism of membrane remodeling by Sar1 remain unresolved. Here, we show that Sar1 transforms synthetic liposomes into structures of different morphologies including tubules and detached vesicles. We demonstrate that Sar1 alone is competent for vesicle scission in a manner that depends on the concentration of Sar1 molecules occupying the membrane. Sar1 molecules align on low-curvature membranes to form an extended lattice. The continuity of this lattice breaks down as the curvature locally increases. The smallest repeating unit constituting the ordered lattice is a Sar1 dimer. The three-dimensional structure of the Sar1 lattice was reconstructed by substituting spherical liposomes with galactoceramide lipid tubules of homogeneous diameter. These data suggest that Sar1 dimerization is responsible for the formation of constrictive membrane curvature. We propose a model whereby Sar1 dimers assemble into ordered arrays to promote membrane constriction and COPII-directed vesicle scission.
已知小GTPase蛋白Sar1参与COPII被膜小泡形成的起始过程以及新生小泡从内质网的脱离过程。Sar1介导膜重塑的分子细节仍未明确。在此,我们表明Sar1可将合成脂质体转变为包括细管和游离小泡在内的不同形态结构。我们证明,Sar1自身就能以一种依赖于占据膜的Sar1分子浓度的方式进行小泡切割。Sar1分子在低曲率膜上排列形成扩展晶格。随着局部曲率增加,这种晶格的连续性被打破。构成有序晶格的最小重复单元是Sar1二聚体。通过用直径均匀的半乳糖神经酰胺脂质细管替代球形脂质体,重建了Sar1晶格的三维结构。这些数据表明,Sar1二聚化负责形成收缩性膜曲率。我们提出一个模型,即Sar1二聚体组装成有序阵列以促进膜收缩和COPII介导的小泡切割。