Retrieval and reuse of pituitary secretory granule proteins.
The pituitary contains professional secretory cells, devoting a large fraction of their energy to the synthesis of hormones that are stored for secretion in response to a complex mixture of inputs. Ba2+, a substitute for Ca2+, and phorbol ester, a mimic for diacylglycerol, have a synergistic effect on exocytosis. By using these secretagogues, we developed a paradigm in which phorbol ester potentiation of Ba2+-evoked exocytosis produces a robust secretory response in multiple pituitary cell types. Because cells subjected to this stimulatory paradigm remain healthy despite their greatly reduced hormone content, we used this paradigm to study the fate of granule membrane proteins. We examined the turnover of peptidylglycine alpha-amidating monooxygenase (PAM), a membrane enzyme involved in the final maturation of many peptides, and VAMP2, a vesicle soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE). The stability of recently synthesized PAM was increased by sustained exocytosis. Biotinylation studies established that the appearance of integral membrane PAM at the plasma membrane was stimulated along with hormone secretion. PAM biotinylated on the cell surface undergoes cleavage to yield soluble peptidylglycine-alpha-hydroxylating monooxygenase that can then be secreted in a regulated fashion. Consistent with a kiss-and-run or cavicapture mode of secretion (Taraska, J. W., Perrais, D., Ohara-Imaizumi, M., Nagamatsu, S., and Almers, W. (2003) Proc. Natl. Acad. Sci. U. S. A. 100, 2070-2075), biotinylated prolactin was also retained by the cells and later released in response to secretagogues. Thus, pituitary cells can retrieve and reuse components of the machinery involved in the final stages of exocytosis (the SNAREs) as well as soluble and membrane granule proteins.
Ferraro
Francesco
F
Neuroscience Department, University of Connecticut Health Center, Farmington, Connecticut 06030-3401, USA.
Eipper
Betty A
BA
Mains
Richard E
RE
D000818:Animals; D001464:Barium / Q000494:pharmacology; D019921:Biotinylation; D002273:Carcinogens / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D004357:Drug Synergism; D005089:Exocytosis / Q000187:drug effects / Q000502:physiology*; D005260:Female; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D009097:Multienzyme Complexes / Q000378:metabolism*; D010903:Pituitary Gland, Anterior / Q000166:cytology / Q000378:metabolism*; D011388:Prolactin / Q000378:metabolism; D050683:R-SNARE Proteins; D051381:Rats; D017207:Rats, Sprague-Dawley; D022142:Secretory Vesicles / Q000187:drug effects / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D002273:Carcinogens; D008565:Membrane Proteins; D009097:Multienzyme Complexes; D050683:R-SNARE Proteins; D001464:Barium; D011388:Prolactin; D006899:Mixed Function Oxygenases; C035189:peptidylglycine monooxygenase; D013755:Tetradecanoylphorbol Acetate