Figure 2.

Immunodetection of dynamin and synaptophysin in chromaffin granules. A, Fractions 2–16 collected from a continuous sucrose density gradient (1.0–2.2 m sucrose) layered with protein extracts from chromaffin cells were subjected to gel electrophoresis and immunodetected with anti-Dyn, anti-DBH, and anti-Syn. B, Purified chromaffin granule membranes were subjected to immunoblotting with anti-Dyn, anti-DBH, and anti-Syn. C, GST alone or GST-Cterm were immobilized on glutathione-agarose beads and incubated with protein extracts from chromaffin granules. The bound material was subjected to immunobloting with anti-Dyn and anti-GST antibodies. D, Chromaffin granule extracts were subjected to immunoprecipitation with anti-Syn. Bound proteins were analyzed by immunoblotting with anti-Dyn and anti-Syn. E, Supernatants from pull-down assays were subjected to immunoprecipitation with anti-Syn, and then bound proteins were analyzed by immunoblotting with anti-Dyn and anti-Syn.