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Although NSF has been proposed to act before docking in yeast vacuolar fusion (24), it is possible that the "predocking" reaction monitored in such in vitro studies is equivalent to the priming observed here and elsewhere (23, 29). For example, docking of synaptic vesicles might lead to a biochemically labile vesicle-membrane interaction that is consolidated during subsequent NSF-regulated priming.
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We thank L. Hawkey and S. Huber for electron microscopy, S. Hilfiker for help with uptake blocker experiments, M. Thomas for use of a flash lamp, A. Niehuis for technical assistance, J. Battey for the squid cDNA library, J. E. Rothman for expression vectors for α-SNAP and NSF, A. Makusky and L. Bonewald for peptide synthesis, and T. Blanpied for critical reading of the manuscript. Supported by grants from NIH, Deutsche Forschungsgemeinschaft, Verband der Chemischen Industrie, and the Human Frontier Science Program.
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