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The Journal of Neuroscience, February 1, 1999, 19(3):900-905
Ca2+-Induced Deprotonation of Peptide Hormones Inside Secretory Vesicles in Preparation for Release
Weiping Han, Danqing Li, Amy K. Stout, Koichi Takimoto, and Edwin S. Levitan Department of Pharmacology, University of Pittsburgh, Pittsburgh, Pennsylvania 15261
The acidic environment inside secretory vesicles ensures that neuropeptides and peptide hormones are packaged in a concentrated condensed form. Although this is optimal for storage, decondensation limits release. Thus, it would be advantageous to alter the physical state of peptides in preparation for exocytosis. Here, we report that depolarization of the plasma membrane rapidly increases enhanced green fluorescent protein (EGFP)-tagged hormone fluorescence inside secretory vesicles. This effect requires Ca2+ influx and persists when exocytosis is inhibited by N-ethylmaleimide. Peptide deprotonation appears to produce this response, because it is not seen when the vesicle pH gradient is collapsed or when a pH-insensitive GFP variant is used. These data demonstrate that Ca2+ evokes alkalinization of the inside of secretory vesicles before exocytosis. Thus, Ca2+ influx into the cytoplasm alters the physical state of intravesicular contents in preparation for release.
Key words: GFP; peptide hormone; exocytosis; alkalinization; secretory vesicle; Ca2+
Copyright © 1999 Society for Neuroscience 0270-6474/99/193900-06$05.00/0
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