Formation of a yeast SNARE complex is accompanied by significant structural changes

FEBS Letters

Rice, Luke M²  Brennwald, Patrick¹  Brünger, Axel T² 

[1] Department of Cell Biology and Anatomy Cornell University Medical College 1300 York Avenue, New York, NY 10021, USA;The Howard Hughes Medical Institute, and Department of Molecular Biophysics and Biochemistry Yale University, New Haven, CT 06520, USA

关键词: Circular dichroism;    Fusion;    Exocytosis;    Yeast;    Structural change;    CD;    circular dichroism;    NSF;    N-ethylmelamide sensitive factor;    SNAP;    soluble NSF attachment protein;    SNARE;    SNAP receptor;    NMR;    nuclear magnetic resonance;    DTT;    dithiothreitol;    SNAP-25;    synaptosomal associated protein of 25 kDa;    VAMP;    vesicle associated membrane protein;    IPTG;    isopropylthiogalactoside;    GST;    glutathione-S-transferase;    SDS;    sodium dodecyl sulphate;    EDTA;    ethylenedinitrilo tetraacetic acid;    PMSF;    phenylmethylsulphonylfluoride;    PCR;    polymerase chain reaction;   

DOI  :  10.1016/S0014-5793(97)01091-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The evolutionarily conserved SNARE (SNAP receptor) proteins and their complexes are key players in the docking and fusion of secretory vesicles with their target membrane. Biophysical techniques were used to characterize structural and energetic properties of the cytoplasmic domains of the yeast SNAREs Snc1 and Sso1, of the SNAP-25-like domain of Sec9, and of the Sso1:Sec9 and Sso1:Sec9:Snc1 complexes. Individually, all three SNAREs are monomeric; Sso1 shows significant secondary structure while Snc1 and Sec9 are largely unstructured. Ternary SNARE complex formation (K _D<50 nM) is accompanied by a more than two-fold increase in secondary structure. This binding induced structure, the large increase in thermal stability, and the self-association of the ternary complex represent conserved properties of SNAREs that are probably important in vesicle docking and fusion.

【 授权许可】

Unknown