title: The Kex2p proregion is essential for the biosynthesis of an active enzyme and requires a C-terminal basic residue for its function.
abstract: The Saccharomyces cerevisiae prohormone-processing enzyme Kex2p is biosynthesized as an inactive precursor extended by its N-terminal proregion. Here we show that deletion of the proregion renders Kex2p inactive both in vivo and in vitro. Absence of the proregion impaired glycosylation and stability and resulted in the retention of the enzyme in the endoplasmic reticulum. These phenotypes were partially complemented by expression of the proregion in trans. Trans complementation was specific to Kex2p proregion because expression of any of the seven mammalian prohormone convertase propeptides had no effect. These data are consistent with a model whereby Kex2p proregion functions as an intramolecular chaperone and indicate that covalent linkage to the protein is not an absolute requirement for proregion function. Furthermore, extensive mutagenesis revealed that, in addition to their function as proteolytic recognition sites, C-terminal basic residues play an active role in proregion-dependent Kex2p activation.
authors.affiliation: Département de Biochimie, Université de Montréal, Montréal, Quebec H3C 3J7, Canada.
authors.forename: G
authors.initials: G
authors.lastname: Lesage
authors.forename: A
authors.initials: A
authors.lastname: Prat
authors.forename: J
authors.initials: J
authors.lastname: Lacombe
authors.forename: D Y
authors.initials: DY
authors.lastname: Thomas
authors.forename: N G
authors.initials: NG
authors.lastname: Seidah
authors.forename: G
authors.initials: G
authors.lastname: Boileau
mesh_terms: D000595:Amino Acid Sequence; D004721:Endoplasmic Reticulum / Q000201:enzymology; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D006031:Glycosylation; D008969:Molecular Sequence Data; D016296:Mutagenesis; D043484:Proprotein Convertases; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins; D013381:Subtilisins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism* / Q000502:physiology
chemical_list: D004792:Enzyme Precursors; D029701:Saccharomyces cerevisiae Proteins; D043484:Proprotein Convertases; D013381:Subtilisins; C058751:KEX2 protein, S cerevisiae