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Mechanistic insights into the recycling machine of the SNARE complex

机译:对SNARE大楼的回收机器的机械洞察力

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摘要

在包括神经传输物释放、荷尔蒙释放和囊泡转运在内的各种细胞过程中,演化上保守的SNARE蛋白形成一个复合物,驱动两个细胞腔的膜之间的融合。融合一旦发生,这些复合物就会被ATP酶NSF和SNAP衔接蛋白拆解,为下一轮的膜融合来循环各个SNARE蛋白。这项研究采用单粒子低温电子显微镜来确定NSF和20S复合物从亚纳米到接近原子分辨率的结构。这篇论文报告了全长度NSF在与ATP和ADP结合状态的结构以及大约660千道尔顿、涉及两个不同SNARE复合物的NSF/SNAP/SNARE(20S)超级复合物的结构。作者的数据为这些重要分子机器的内部工作方式提供了前所未有的详细信息。%Evolutionarily conserved SNARE (soluble N-ethylmaleimide sensitive factor attachment protein receptors) proteins form a complex that drives membrane fusion in eukaryotes. The ATPase NSF (N-ethylmaleimide sensitive factor), together with SNAPs (soluble NSF attachment protein), disassembles the SNARE complex into its protein components, making individual SNAREs available for subsequent rounds of fusion. Here we report structures of ATP- and ADP-bound NSF, and the NSF/ SNAP/SNARE (20S) supercomplex determined by single-particle electron cryomicroscopy at near-atomic to sub -nanometre resolution without imposing symmetry. Large, potentially force-generating, conformational differences exist between ATP- and ADP-bound NSF. The 20S supercomplex exhibits broken symmetry, transitioning from six-fold symmetry of the NSF ATPase domains to pseudo four-fold symmetry of the SNARE complex. SNAPs interact with the SNARE complex with an opposite structural twist, suggesting an unwinding mechanism. The interfaces between NSF, SNAPs, and SNAREs exhibit characteristic electrostatic patterns, suggesting how one NSF/SNAP species can act on many different SNARE complexes.
机译:在包括神经传输物释放、荷尔蒙释放和囊泡转运在内的各种细胞过程中,演化上保守的SNARE蛋白形成一个复合物,驱动两个细胞腔的膜之间的融合。融合一旦发生,这些复合物就会被ATP酶NSF和SNAP衔接蛋白拆解,为下一轮的膜融合来循环各个SNARE蛋白。这项研究采用单粒子低温电子显微镜来确定NSF和20S复合物从亚纳米到接近原子分辨率的结构。这篇论文报告了全长度NSF在与ATP和ADP结合状态的结构以及大约660千道尔顿、涉及两个不同SNARE复合物的NSF/SNAP/SNARE(20S)超级复合物的结构。作者的数据为这些重要分子机器的内部工作方式提供了前所未有的详细信息。%Evolutionarily conserved SNARE (soluble N-ethylmaleimide sensitive factor attachment protein receptors) proteins form a complex that drives membrane fusion in eukaryotes. The ATPase NSF (N-ethylmaleimide sensitive factor), together with SNAPs (soluble NSF attachment protein), disassembles the SNARE complex into its protein components, making individual SNAREs available for subsequent rounds of fusion. Here we report structures of ATP- and ADP-bound NSF, and the NSF/ SNAP/SNARE (20S) supercomplex determined by single-particle electron cryomicroscopy at near-atomic to sub -nanometre resolution without imposing symmetry. Large, potentially force-generating, conformational differences exist between ATP- and ADP-bound NSF. The 20S supercomplex exhibits broken symmetry, transitioning from six-fold symmetry of the NSF ATPase domains to pseudo four-fold symmetry of the SNARE complex. SNAPs interact with the SNARE complex with an opposite structural twist, suggesting an unwinding mechanism. The interfaces between NSF, SNAPs, and SNAREs exhibit characteristic electrostatic patterns, suggesting how one NSF/SNAP species can act on many different SNARE complexes.

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  • 来源
    《Nature》 |2015年第7537期|61-67a1|共8页
  • 作者单位

    Department of Molecular and Cellular Physiology, Howard Hughes Medical Institute, Stanford University, Stanford, California 94305, USA;

    Keck Advanced Microscopy Laboratory, Department of Biochemistry and Biophysics, University of California, San Francisco, California 94158, USA;

    Department of Molecular and Cellular Physiology, Howard Hughes Medical Institute, Stanford University, Stanford, California 94305, USA;

    Department of Molecular and Cellular Physiology, Howard Hughes Medical Institute, Stanford University, Stanford, California 94305, USA;

    Department of Molecular and Cellular Physiology, Howard Hughes Medical Institute, Stanford University, Stanford, California 94305, USA;

    Keck Advanced Microscopy Laboratory, Department of Biochemistry and Biophysics, University of California, San Francisco, California 94158, USA;

    Department of Molecular and Cellular Physiology, Howard Hughes Medical Institute, Stanford University, Stanford, California 94305, USA,Department of Neurology and Neurological Sciences, Department of Structural Biology, Department of Photon Science, Stanford University, Stanford, California 94305, USA;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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  • 正文语种 eng
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