Electrostatic interactions between the syntaxin membrane anchor and neurotransmitter passing through the fusion pore.
Recent experiments have shown that flux through the fusion pore is sensitive to manipulations of the side-chain size of certain residues in the syntaxin (syx) membrane anchor. These residues were proposed to line the wall of the fusion pore of Ca(2+)-triggered exocytosis. Here we continued this line of experimentation to examine possible electrostatic interactions between the pore lining residues and the neurotransmitter norepinephrine (NE). Replacing syx pore-lining residues with aspartate enhanced NE flux above that expected for the size of the aspartate side chain. In contrast, substitution with arginine reduced NE flux below that expected for the size of its side chain. Substituting aspartate and arginine into the nonpore-lining residues did not alter the fusion pore flux. Other amino acids with ionizable side chains had variable effects. These results indicate an electrostatic interaction between the pore-lining residues of syx and NE, and provide additional evidence that the syx membrane anchor is a structural component of the fusion pore.
Han
Xue
X
Department of Physiology, University of Wisconsin, Madison, Wisconsin 53706, USA.
Jackson
Meyer B
MB
D019943:Amino Acid Substitution; D000818:Animals; D001693:Biological Transport, Active / Q000502:physiology; D002463:Cell Membrane Permeability / Q000502:physiology*; D005089:Exocytosis / Q000502:physiology; D008561:Membrane Fusion / Q000502:physiology; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D018377:Neurotransmitter Agents / Q000737:chemistry* / Q000378:metabolism*; D009638:Norepinephrine / Q000737:chemistry* / Q000378:metabolism*; D016716:PC12 Cells; D016062:Porosity; D050765:Qa-SNARE Proteins; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D055672:Static Electricity; D013329:Structure-Activity Relationship
D008565:Membrane Proteins; D018377:Neurotransmitter Agents; D050765:Qa-SNARE Proteins; D011994:Recombinant Proteins; D009638:Norepinephrine