title: Chaperone interactions of the metalloproteinase meprin A in the secretory or proteasomal-degradative pathway.
abstract: The secreted form of mouse meprin A is a homooligomer of meprin alpha subunits that contain a prosequence, a catalytic domain, and three domains designated as MAM (meprin, A5 protein, receptor protein-tyrosine phosphatase mu), MATH (meprin and TRAF homology), and AM (AfterMath). Previous studies indicated that wild-type mouse meprin alpha is predominantly a secreted protein, while the MAM deletion mutant (DeltaMAM) is degraded intracellularly. The work herein indicates that the DeltaMAM mutant is ubiquitinated and degraded via the proteasomal pathway. Both wild-type meprin alpha and the DeltaMAM mutant interact with the molecular chaperones calnexin and calreticulin in the endoplasmic reticulum. The interactions of the chaperones with the DeltaMAM mutant were significantly prolonged in the presence of lactacystin, a specific inhibitor of the proteasome, whereas those with the wild type were not affected by this inhibitor. Trimming of the Asn-linked core oligosaccharides of meprin subunits was required for interactions with the chaperones. The data indicated that folding of the wild-type protein was accelerated by chaperones, whereas the rate of dimerization was unaffected. Thus, calnexin and calreticulin are intimately involved in the correct folding and transport of meprin to the plasma membrane, as well as in retrograde transport of the DeltaMAM mutant to the ubiquitin-dependent proteasomal degradative pathway in the cytosol.
authors.affiliation: Department of Biochemistry and Molecular Biology, The Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033-0850, USA.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Tsukuba
authors.forename: Tomoko
authors.initials: T
authors.lastname: Kadowaki
authors.forename: Jeremy A
authors.initials: JA
authors.lastname: Hengst
authors.forename: Judith S
authors.initials: JS
authors.lastname: Bond
mesh_terms: D000111:Acetylcysteine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D002135:Calcium-Binding Proteins / Q000378:metabolism; D037281:Calnexin; D037282:Calreticulin; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism; D007212:Indolizines / Q000494:pharmacology; D008666:Metalloendopeptidases / Q000378:metabolism*; D051379:Mice; D008954:Models, Biological; D018832:Molecular Chaperones / Q000378:metabolism*; D009097:Multienzyme Complexes / Q000378:metabolism*; D009154:Mutation; D009844:Oligosaccharides / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex; D011498:Protein Precursors / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D012261:Ribonucleoproteins / Q000378:metabolism; D017384:Sequence Deletion; D025801:Ubiquitin / Q000378:metabolism; D054836:Indolizidines
chemical_list: D000111:Acetylcysteine; D002135:Calcium-Binding Proteins; D037281:Calnexin; D037282:Calreticulin; D003546:Cysteine Endopeptidases; D015853:Cysteine Proteinase Inhibitors; D006023:Glycoproteins; D007212:Indolizines; D008666:Metalloendopeptidases; D018832:Molecular Chaperones; D009097:Multienzyme Complexes; D009844:Oligosaccharides; D046988:Proteasome Endopeptidase Complex; D011498:Protein Precursors; D012261:Ribonucleoproteins; D025801:Ubiquitin; C067713:lactacystin; C053833:meprin A; C037806:castanospermine; D054836:Indolizidines