Thiol and aspartyl proteolytic activities in secretory vesicles of bovine pituitary.
Thiol and aspartyl proteolytic activities in isolated secretory vesicles of neural (NL) and intermediate (IL) lobes of bovine pituitary were characterized with heterologous enkephalin and tachykinin precursor substrates, 35S-(Met)-preproenkephalin and 35S-(Met)-beta-preprotachykinin. IL and NL secretory vesicles contained thiol-dependent proteolytic activity that cleaved the enkephalin precursor with a pH optimum of 4.5; this activity resembled a novel "prohormone thiol protease' previously purified and characterized from adrenal medulla chromaffin granules. IL and NL vesicles also demonstrated aspartyl proteolytic activity with acidic pH optimum, as shown by pepstatin A inhibition of tachykinin and enkephalin precursor cleaving activity. This activity may be related to a previously characterized chromaffin granule aspartyl protease (CGAP) related to cathepsin D (2), as indicated by the presence of immunoreactive CGAP in NL secretory vesicles by anti-CGAP immunoblots. These results show that pituitary secretory vesicles, like chromaffin granules, may contain similar thiol-dependent and aspartyl proteolytic activities.
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Department of Biochemistry, Uniformed Services University of the Health Sciences, Bethesda, MD 20814.
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D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000187:drug effects / Q000378:metabolism*; D002417:Cattle; D002837:Chromaffin Granules / Q000201:enzymology; D003546:Cysteine Endopeptidases / Q000187:drug effects / Q000378:metabolism*; D003594:Cytoplasmic Granules / Q000201:enzymology*; D004745:Enkephalins / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D010902:Pituitary Gland / Q000201:enzymology* / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology; D011498:Protein Precursors / Q000378:metabolism; D015320:Tachykinins / Q000378:metabolism
D004745:Enkephalins; D011480:Protease Inhibitors; D011498:Protein Precursors; D015320:Tachykinins; C044206:preprotachykinin; C033660:preproenkephalin; D003546:Cysteine Endopeptidases; D016282:Aspartic Acid Endopeptidases