Insertion of the membrane-proximal region of the neuronal SNARE coiled coil into the membrane.
In the neuron, soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins assemble into an alpha-helical coiled coil that bridges the synaptic vesicle to the plasma membrane and drives membrane fusion, a required process for neurotransmitter release at the nerve terminal. How does coiled coil formation drive membrane fusion? To investigate the structural and energetic coupling between the coiled coil and membrane, the recombinant SNARE complex in the phospholipid bilayer was studied using fluorescence quenching and site-directed spin labeling EPR. Fluorescence analysis revealed that two native Trp residues at the membrane-proximal region of the coiled coil are inserted into the membrane, tightly coupling the coiled coil to the membrane. The EPR results indicate that the coiled coil penetrates into the membrane with an oblique angle, providing a favorable geometry for the basic residues to interact with negatively charged lipids. The result supports the proposition that core complex formation directly leads to the apposition of two membranes, which could facilitate lipid mixing. Trp residues and basic residues are abundant at the membrane-proximal region of transmembrane SNARE proteins, suggesting the generality of the proposed mechanism for the SNARE complex-membrane coupling.
Kweon
Dae-Hyuk
DH
Department of Biochemistry, Biophysics, and Molecular Biology, Iowa State University, Ames, Iowa 50011, USA.
Kim
Chang Sup
CS
Shin
Yeon-Kyun
YK
D004578:Electron Spin Resonance Spectroscopy; D008561:Membrane Fusion / Q000502:physiology*; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics*; D016254:Mutagenesis, Insertional; D017433:Protein Structure, Secondary; D050683:R-SNARE Proteins; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D050600:SNARE Proteins; D013113:Spin Labels; D033921:Vesicular Transport Proteins
D008565:Membrane Proteins; D050683:R-SNARE Proteins; D011994:Recombinant Proteins; D050600:SNARE Proteins; D013113:Spin Labels; D033921:Vesicular Transport Proteins