Targeting of membrane proteins to the regulated secretory pathway in anterior pituitary endocrine cells.
Unlike the neuroendocrine cell lines widely used to study trafficking of soluble and membrane proteins to secretory granules, the endocrine cells of the anterior pituitary are highly specialized for the production of mature secretory granules. Therefore, we investigated the trafficking of three membrane proteins in primary anterior pituitary endocrine cells. Peptidylglycine alpha-amidating monooxygenase (PAM), an integral membrane protein essential to the production of many bioactive peptides, is cleaved and enters the regulated secretory pathway even when expressed at levels 40-fold higher than endogenous levels. Myc-TMD/CD, a membrane protein lacking the lumenal, catalytic domains of PAM, is still stored in granules. Secretory granules are not the default pathway for all membrane proteins, because Tac accumulates on the surface of pituitary endocrine cells. Overexpression of PAM is accompanied by a diminution in its endoproteolytic cleavage and in its BaCl(2)-stimulated release from mature granules. Because internalized PAM/PAM-antibody complexes are returned to secretory granules, the endocytic machinery of the pituitary endocrine cells is not saturated. As in corticotrope tumor cells, expression of PAM or Myc-TMD/CD alters the organization of the actin cytoskeleton. PAM-mediated alterations in the cytoskeleton may limit maturation of PAM and storage in mature granules.
El Meskini
R
R
Department of Neuroscience, University of Connecticut Health Center, Farmington, Connecticut 06030-3401, USA.
Galano
G J
GJ
Marx
R
R
Mains
R E
RE
Eipper
B A
BA
D000199:Actins / Q000502:physiology; D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D004702:Endocrine Glands / Q000166:cytology / Q000378:metabolism* / Q000502:physiology; D004705:Endocytosis / Q000502:physiology; D066298:In Vitro Techniques; D008297:Male; D008565:Membrane Proteins / Q000187:drug effects / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000187:drug effects / Q000378:metabolism*; D009097:Multienzyme Complexes; D010903:Pituitary Gland, Anterior / Q000166:cytology / Q000378:metabolism* / Q000502:physiology; D010908:Pituitary Hormones, Anterior / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013347:Subcellular Fractions; D014162:Transfection
D000199:Actins; D008565:Membrane Proteins; D009097:Multienzyme Complexes; D010908:Pituitary Hormones, Anterior; D016271:Proto-Oncogene Proteins c-myc; D006899:Mixed Function Oxygenases; C035189:peptidylglycine monooxygenase