title: Binding of BiP to the processing enzyme lymphoma proprotein convertase prevents aggregation, but slows down maturation.
abstract: Lymphoma proprotein convertase (LPC) is a subtilisin-like serine protease of the mammalian proprotein convertase family. It is synthesized as an inactive precursor protein, and propeptide cleavage occurs via intramolecular cleavage in the endoplasmic reticulum. In contrast to other convertases like furin and proprotein convertase-1, propeptide cleavage occurs slowly. Also, both a glycosylated and an unglycosylated precursor are detected. Here we demonstrate that the unglycosylated precursor form of LPC is localized in the cytosol due to the absence of a signal peptide. Using a reducible cross-linker, we found that glycosylated pro-LPC is associated with the molecular chaperone BiP. In addition, we show that pro-LPC is prone to aggregation and forms large complexes linked via interchain disulfide bonds. BiP is associated mainly with non-aggregated pro-LPC and pro-LPC dimers and trimers, suggesting that BiP prevents aggregation. Overexpression of wild-type BiP or a dominant-negative BiP ATPase mutant resulted in reduced processing of pro-LPC. Taken together, these results suggest that binding of BiP to pro-LPC prevents aggregation, but results in slower maturation.
authors.affiliation: Laboratory for Molecular Oncology, Center for Human Genetics, University of Leuven and Flanders Interuniversity Institute for Biotechnology, Leuven, Belgium. john.creemers@med.kuleuven.ac.be
authors.forename: J W
authors.initials: JW
authors.lastname: Creemers
authors.forename: J W
authors.initials: JW
authors.lastname: van de Loo
authors.forename: E
authors.initials: E
authors.lastname: Plets
authors.forename: L M
authors.initials: LM
authors.lastname: Hendershot
authors.forename: W J
authors.initials: WJ
authors.lastname: Van De Ven
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D019943:Amino Acid Substitution; D000818:Animals; D001665:Binding Sites; D016466:CHO Cells; D019556:COS Cells; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002522:Chlorocebus aethiops; D006224:Cricetinae; D000091342:Endoplasmic Reticulum Chaperone BiP; D006031:Glycosylation; D006360:Heat-Shock Proteins; D008322:Mammals; D018832:Molecular Chaperones / Q000302:isolation & purification / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D012694:Serine; D012697:Serine Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D000091342:Endoplasmic Reticulum Chaperone BiP; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D011994:Recombinant Proteins; D012694:Serine; D012697:Serine Endopeptidases; D000251:Adenosine Triphosphatases