Acquisition of Lubrol insolubility, a common step for growth hormone and prolactin in the secretory pathway of neuroendocrine cells.
Rat prolactin in the dense cores of secretory granules of the pituitary gland is a Lubrol-insoluble aggregate. In GH(4)C(1) cells, newly synthesized rat prolactin and growth hormone were soluble, but after 30 min about 40% converted to a Lubrol-insoluble form. Transport from the endoplasmic reticulum is necessary for conversion to Lubrol insolubility, since incubating cells with brefeldin A or at 15 degrees C reduced formation of insoluble rat (35)S-prolactin. Formation of Lubrol-insoluble aggregates has protein and cell specificity; newly synthesized human growth hormone expressed in AtT20 cells underwent a 40% conversion to Lubrol insolubility with time, but albumin did not, and human growth hormone expressed in COS cells underwent less than 10% conversion to Lubrol insolubility. del32-46 growth hormone, a naturally occurring form of growth hormone, and P89L growth hormone underwent conversion, although they were secreted more slowly, indicating that there is some tolerance in structural requirements for aggregation. An intracellular compartment with an acidic pH is not necessary for conversion to Lubrol insolubility, because incubation with chloroquine or bafilomycin slowed, but did not prevent, the conversion. GH(4)C(1) cells treated with estradiol, insulin, and epidermal growth factor accumulate more secretory granules and store more prolactin, but not more growth hormone, than untreated cells; Lubrol-insoluble aggregates of prolactin and growth hormone formed to the same extent in hormone-treated or untreated GH(4)C(1) cells, but prolactin was retained longer in hormone-treated cells. These findings indicate that aggregation alone is not sufficient to cause retention of secretory granule proteins, and there is an additional selective process.
Lee
M S
MS
Department of Pharmacology, Yale University School of Medicine, New Haven, Connecticut 06520, USA.
Zhu
Y L
YL
Chang
J E
JE
Dannies
P S
PS
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D020126:Brefeldin A / Q000494:pharmacology; D019556:COS Cells; D002738:Chloroquine / Q000494:pharmacology; D004136:Dinitrobenzenes / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D004815:Epidermal Growth Factor / Q000494:pharmacology; D004958:Estradiol / Q000494:pharmacology; D019382:Human Growth Hormone / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007328:Insulin / Q000494:pharmacology; D018942:Macrolides; D009154:Mutation; D010902:Pituitary Gland / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000494:pharmacology*; D011388:Prolactin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport / Q000187:drug effects; D051381:Rats; D022142:Secretory Vesicles / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D012995:Solubility / Q000187:drug effects; D013379:Substrate Specificity; D014407:Tumor Cells, Cultured; D014461:Ultracentrifugation
D000900:Anti-Bacterial Agents; D004136:Dinitrobenzenes; D007328:Insulin; D018942:Macrolides; C057620:bafilomycin A; D019382:Human Growth Hormone; D020126:Brefeldin A; D011092:Polyethylene Glycols; D004958:Estradiol; C005092:lubrol; D004815:Epidermal Growth Factor; D002738:Chloroquine; D011388:Prolactin; C042432:3-(2,4-dinitroanilino)-3'-amino-N-methyldipropylamine