Sorting of PC2 to the regulated secretory pathway in AtT20 cells.
PC2 and PC3 are neuroendocrine specific members of the eukaryotic subtilisin-like proprotein convertase (PC) family. Both are sorted via the regulated secretory pathway into secretory granules. In order to identify sequences in PC2 which are involved in targeting to the regulated secretory pathway we expressed a series of PC2 cDNAs containing mutations in the C terminal or propeptide domains in the mouse corticotrophic AtT20 cell line. Sorting of endogenous PC3 was used as a control. PC2 and PC3 were secreted with similar kinetics and sorted to secretory granules with similar efficiencies. Deletions of up to 50 amino acids from the C-terminus of proPC2 had no effect on secretion or sorting, but larger deletions completely prevented maturation or secretion. Two large deletions within the propeptide also prevented secretion. Smaller deletions between the primary and secondary cleavage sites, or of the primary cleavage site, reduced the amount of protein secreted but did not affect sorting to secretory granules. Replacement of the propeptide of PC2 with that of the endogenous PC3 also had no effect on secretion or sorting. The results indicate that targeting of proPC2 to the regulated secretory pathway is dependent on more than one region within the proPC2 molecule.
Taylor
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NA
Department of Molecular and Cell Biology, University of Aberdeen, Institute of Medical Sciences, Foresterhill, UK.
Jan
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Scougall
K T
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Docherty
K
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Shennan
K I
KI
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D004792:Enzyme Precursors / Q000378:metabolism; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D016296:Mutagenesis; D045706:Proprotein Convertase 2; D011499:Protein Processing, Post-Translational; D017386:Sequence Homology, Amino Acid; D013381:Subtilisins / Q000737:chemistry / Q000378:metabolism*
D004792:Enzyme Precursors; D013381:Subtilisins; D045706:Proprotein Convertase 2