Identification of d-amino acid oxidase and propiverine interaction partners and their potential role in the propiverine-mediated nephropathy. (1st February 2018)

• Record Type: Journal Article
• Authors: Maier, Marcia Y.; Luks, Lisanne; Baudendistel, Oliver R.; Wittmann, Valentin; Dietrich, Daniel R.
• Is Part Of: Chemico-biological interactions. Volume 281(2018)
• Page Start: 69
• Page End: 80
• Publication Date: 2018-02-01
• Subjects: d-amino acid oxidase (DAAO) -- Propiverine -- Peroxisomal import -- Protein trafficking -- Cytosolic and nuclear protein mislocalization and accumulation -- Co-immunoprecipitation (Co-IP) -- Nano-LC-ESI-MS/MS -- Immunohistochemistry (IHC); hDAAO, rDAAO human and rat d-amino acid oxidase -- PTS1 peroxisomal targeting signal 1 -- PTS2 peroxisomal targeting signal 2 -- PMPs peroxisomal membrane proteins -- PEX peroxin -- HEK293 human embryonic kidney -- WKPT Wistar-Kyoto rat kidney proximal tubule; Biochemistry -- Periodicals; Toxicological chemistry -- Periodicals; Biochemistry -- Periodicals; Biologie moléculaire -- Périodiques; Biochimie -- Périodiques; Toxicologie biochimique -- Périodiques; 572
• DOI: 10.1016/j.cbi.2017.12.023
• Languages: English
• ISSNs: 0009-2797

Abstract

Abstract: Propiverine, a frequently-prescribed pharmaceutical for the treatment of symptoms associated with overactive bladder syndrome, provoked massive intranuclear and cytosolic protein inclusions in rat proximal tubule epithelium, primarily consisting of the peroxisomal targeting signal 1 (PTS1) containing proteind -amino acid oxidase (DAAO). As this type of nephropathy was also observed for other drugs, the aim was to determine whether propiverine interferes with trafficking and/or import of peroxisomal proteins. To elucidate this, DAAO- and propiverine-specific interaction partners from human HEK293 and rat WKPT cell lines and rat kidney and liver homogenate were determined using co-immunoprecipitation with subsequent nano-ESI-LC-MS/MS analyses. Corroboration of the role of DAAO- and/or propiverine-specific interaction partners in the drug-induced DAAO accumulation was sought via specific immunofluorescence staining of rat kidney sections from control and propiverine-treated rats. Above analyses demonstrated the interaction of propiverine with several protein classes, foremost peroxisomal proteins (DAAO, MFE2, HAOX2) and proteins of the protein quality control system, i.e. chaperones (HSP70 and DnaJ co-chaperones), proteases and proteasomal proteins (regulatory subunits of the 26S proteasome; Rpn1/2). The immunofluorescence analysis revealed mislocalization of many PTS1-proteins (DAAO, CAT, MFE2, ACOX1, EHHADH) in rat renal sections, strongly suggesting that propiverine primarily binds to PTS1 proteins resulting in the formation of PTS1 but not PTS2 or peroxisomal membrane protein (PMP) accumulations. Moreover, chaperones involved in peroxisomal trafficking (HSC70, DnaJB1) and peroxisomal biogenesis factor proteins (PEX3, PEX5, PEX7), also presented with distinct mislocalization patterns. Concomitantly, an increased number of peroxisomes was observed, suggestive of a compensatory mechanism for the presumably suboptimally functioning peroxisomes. Overall, the data presented suggested that propiverine interacts exclusively with DAAO or with a selected number of PTS1 proteins. The consequence of this interaction is the abrogated trafficking and peroxisomal import of PTS1 proteins concomitant with their nuclear and cytosolic accumulation due to inhibited degradation and imbalanced protein homeostasis. Graphical abstract: Propiverine's dual effect on localization and degradation of PTS1 proteins in proximal tubule epithelial cells: Proposed mechanisms. Highlights: d -amino acid oxidase (DAAO) interacts with catalase and MFE2 in vitro . A biotinylated propiverine derivative was efficiently synthesized. Propiverine interacts with PTS1 proteins (DAAO, MFE2, HAOX2) and 26S Rpn1 and Rpn2. Propiverine interacts with oxidoreductases in rat kidney and liver homogenate. Propiverine exposure does not induce accumulation of PTS2 PMPs in rat kidney.