Adenovirally encoded prohormone convertase-1 functions in atrial myocyte large dense core vesicles.
Bioactive peptides are usually synthesized as inactive precursor proteins that yield bioactive products only after specific biosynthetic processing events. Large dense core vesicles (LDCV) are usually the site of storage of mature peptides. Atrial myocyte LDCV are rather unique in their storage of intact prohormone, proatrial natriuretic factor (pro-ANF), with no storage of cleaved products. To investigate whether the lack of intracellular cleavage of pro-ANF is due to the absence of prohormone convertases (PCs) from the atrial granules or to other factors, we expressed PC1 in atrial myocyte cultures using a recombinant adenovirus vector. Pro-PC1 protein was processed to mature PC1 and to the COOH-terminally shortened neuroendocrine-specific form of PC1 and rapidly secreted. Integral membrane forms of peptidylglycine alpha-amidating monooxygenase (PAM) were processed by PC1, and two primary products were secreted: a monofunctional monooxygenase and a larger bifunctional form. The cleaved PAM products were stored in LDCV, as secretion of PAM-derived products was stimulatable. In addition, pro-ANF was processed to ANF within PC1-expressing cells. In primary atrial myocytes, virally encoded PC1 is active on three substrates; lack of cleavage of pro-ANF and PAM in atrial myocytes is not due to a fundamental inability of atrial LDCV to support endoproteolytic processing.
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Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205-2185, USA.
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D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000235:genetics* / Q000378:metabolism*; D009320:Atrial Natriuretic Factor / Q000378:metabolism; D002478:Cells, Cultured; D003594:Cytoplasmic Granules / Q000201:enzymology*; D006325:Heart Atria; D006899:Mixed Function Oxygenases / Q000378:metabolism; D009097:Multienzyme Complexes; D009206:Myocardium / Q000166:cytology / Q000201:enzymology*; D043484:Proprotein Convertases; D011498:Protein Precursors / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D014018:Tissue Distribution
D009097:Multienzyme Complexes; D011498:Protein Precursors; D009320:Atrial Natriuretic Factor; D006899:Mixed Function Oxygenases; C035189:peptidylglycine monooxygenase; D043484:Proprotein Convertases; D016282:Aspartic Acid Endopeptidases