PACE4: a subtilisin-like endoprotease with unique properties.
PACE4 is one of the neuroendocrine-specific mammalian subtilisin-related endoproteases believed to function in the secretory pathway. The biosynthesis and secretion of PACE4 have been studied using transfected neuroendocrine and fibroblast cell lines. as well as primary pituitary cultures. ProPACE4 (approx. 106 kDa) is cleaved intracellularly before secretion of PACE4 (approx. 97 kDa); the N-terminal propeptide cleavage is accelerated in a truncated form of PACE4 lacking the Cys-rich C-terminal region (PACE4s). Neither PACE4 nor PACE4s is stored in regulated neuroendocrine secretory granules, whereas pro-opiomelanocortin-derived peptides and prohormone convertase I enter the regulated secretory pathway efficiently. The relatively slow cleavage of the proregion of proPACE4 in primary anterior pituitary cells, followed by rapid secretion of PACE4, is similar to the results for proPACE4 in transfected cell lines. The enzyme activity of PACE4 is distinct from furin and prohormone convertases, both in the marked sensitivity of PACE4 to inhibition by leupeptin and the relative insensitivity of PACE4 to inhibition by Ca2+ chelators and dithiothreitol; PACE4 is not inhibited by the alpha1-antitrypsin Portland variant that is very potent at inhibiting furin. The unique biosynthetic and enzymic patterns seen for PACE4 suggest a role for this neuroendocrine-specific subtilisin-like endoprotease outside the pathway for peptide biosynthesis.
Mains
R E
RE
Department of Neuroscience, The Johns Hopkins University School of Medicine, Baltimore, MD 21205-2185, U.S.A.
Berard
C A
CA
Denault
J B
JB
Zhou
A
A
Johnson
R C
RC
Leduc
R
R
D015153:Blotting, Western; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004720:Endonucleases / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology; D045683:Furin; D015870:Gene Expression / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D010902:Pituitary Gland / Q000201:enzymology*; D043484:Proprotein Convertases; D011499:Protein Processing, Post-Translational / Q000502:physiology; D012697:Serine Endopeptidases / Q000096:biosynthesis / Q000737:chemistry* / Q000378:metabolism; D013381:Subtilisins / Q000037:antagonists & inhibitors / Q000378:metabolism; D014162:Transfection / Q000235:genetics; D001615:beta-Endorphin / Q000494:pharmacology; D008083:beta-Lipotropin / Q000494:pharmacology
D018076:DNA, Complementary; D004791:Enzyme Inhibitors; D001615:beta-Endorphin; D008083:beta-Lipotropin; D004720:Endonucleases; C080654:PCSK6 protein, human; D043484:Proprotein Convertases; D012697:Serine Endopeptidases; D013381:Subtilisins; D045683:Furin