Organization of synaptic junction proteins.
Most of the polypeptides of isolated brain synaptic junction preparations are cross-linked by disulfide bonds; these bonds are readily reformed following reduction by ?-mercaptoethanol, suggesting that other, non-covalent interactions may hold the polypeptides in close contact. When synaptic junctions were treated with ?-mercaptoethanol in conjunction with a large variety of reagents known to disrupt certain types of non-covalent bonds, however, the polypeptides could still be cross-linked. Furthermore, virtually all of these species remained associated with ?-mercaptoethanol-reduced membrane in the presence of 0.05 N NaOH or 0.5% Triton X-100, although each of these solvents extracts large amounts of protein from extra-junctional membrane, which has a composition apparently similar to that of synaptic junctions. These results indicate that any non-covalent interactions existing among synaptic junction proteins are inaccessible to treatments at the surface of the membrane, being located within the lipid bilayer or perhaps at points of contact with sub-membranous arrays such as the post-synaptic density. Under special conditions, the polypeptides of extra-junctional membrane can also be induced to form disulfide cross-links among one another, and these bonds can likewise be rapidly reformed following reduction, giving this membrane properties similar to that of the junction. This suggests a model by which stabilized junctional membrane may be formed from fluid extra-junctional membrane during synaptogenesis.
Smith
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Department of Pharmacology, University of California, San Francisco, San Francisco, CA 94137, U.S.A.
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