Proinsulin endoproteolysis confers enhanced targeting of processed insulin to the regulated secretory pathway.
Recently, two different prohormone-processing enzymes, prohormone convertase 1 (PC1) and carboxypeptidase E, have been implicated in enhancing the storage of peptide hormones in endocrine secretory granules. It is important to know the extent to which such molecules may act as "sorting receptors" to allow the selective trafficking of cargo proteins from the trans-Golgi network into forming granules, versus acting as enzymes that may indirectly facilitate intraluminal storage of processed hormones within maturing granules. GH4C1 cells primarily store prolactin in granules; they lack PC1 and are defective for intragranular storage of transfected proinsulin. However, proinsulin readily enters the immature granules of these cells. Interestingly, GH4C1 clones that stably express modest levels of PC1 store more proinsulin-derived protein in granules. Even in the presence of PC1, a sizable portion of the proinsulin that enters granules goes unprocessed, and this portion largely escapes granule storage. Indeed, all of the increased granule storage can be accounted for by the modest portion converted to insulin. These results are not unique to GH4C1 cells; similar results are obtained upon PC1 expression in PC12 cells as well as in AtT20 cells (in which PC1 is expressed endogenously at higher levels). An in vitro assay of protein solubility indicates a difference in the biophysical behavior of proinsulin and insulin in the PC1 transfectants. We conclude that processing to insulin, facilitated by the catalytic activities of granule proteolytic enzymes, assists in the targeting (storage) of the hormone.
Kuliawat
R
R
Division of Endocrinology, Department of Developmental and Molecular Biology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
Prabakaran
D
D
Arvan
P
P
D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000096:biosynthesis* / Q000235:genetics; D002460:Cell Line; D003594:Cytoplasmic Granules / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D008954:Models, Biological; D016716:PC12 Cells; D011384:Proinsulin / Q000235:genetics / Q000378:metabolism*; D011388:Prolactin / Q000378:metabolism; D043484:Proprotein Convertases; D011499:Protein Processing, Post-Translational; D051381:Rats; D015398:Signal Transduction; D013997:Time Factors; D014162:Transfection
D011388:Prolactin; D011384:Proinsulin; D043484:Proprotein Convertases; D016282:Aspartic Acid Endopeptidases