Post-translational processing and secretory pathway of human atriopeptin in rat pheochromocytoma cells.
Atriopeptin (AP) is expressed in several tissues with each tissue capable of specific differences in processing of the prohormone (pro-AP) to mature low molecular forms of the peptide. Since pro-AP has low biological activity, processing into mature AP is a critical activation event. This observation prompted us to study whether granule storage or regulated secretion of AP is essential for cleavage of mature peptide. We examined the processing of AP in adrenal medulla derived cells, using the rat pheochromocytoma cell line (PC12 cell) stably transfected with a genomic human AP DNA in the presence and absence of nerve growth factor (NGF), and also examined the mechanism of AP secretion and compared the results with those obtained using transfected chinese hamster ovary cells (CHO cells). The amount of prohormone was 5-10 fold higher than that of low molecular form of AP in the transfected PC12 cells. This ratio was essentially unchanged in differentiated PC12 cells after NGF treatment of the cells. Potassium depolarization of the transfected PC12 cells caused a 5-fold increase in AP release into the medium primarily as the intact prohormone. On the other hand, transfected CHO cells only exhibited constitutive AP release which is non-response to depolarization. These results suggest that the AP prohormone is sorted into secretory granules as the prohormone in PC12 cells and undergoes regulated release in response to depolarization indicating granule storage or release is not the critical determinant of AP prohormone cleavage.
Shiono
S
S
Department of Pharmacology, Washington University School of Medicine, St. Louis, Missouri 63110.
Suganuma
N
N
Bo
M
M
Boime
I
I
Seibert
K
K
Nakao
K
K
Mukoyama
M
M
Imura
H
H
Needleman
P
P
D000310:Adrenal Gland Neoplasms; D000818:Animals; D009320:Atrial Natriuretic Factor / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D003412:Cricetulus; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D007700:Kinetics; D009414:Nerve Growth Factors / Q000494:pharmacology; D010053:Ovary; D010673:Pheochromocytoma; D010957:Plasmids; D011189:Potassium Chloride / Q000494:pharmacology; D011498:Protein Precursors / Q000235:genetics*; D011499:Protein Processing, Post-Translational; D011863:Radioimmunoassay; D051381:Rats; D015183:Restriction Mapping; D014162:Transfection
D009414:Nerve Growth Factors; D011498:Protein Precursors; D011189:Potassium Chloride; D009320:Atrial Natriuretic Factor