title: Detection of mutant uromodulin in transgenic mouse harboring a mutant human UMOD gene.
abstract: Uromodulin is the most abundant protein secreted in urine, and the mutated form of the uromodulin gene is associated with uromodulin-associated kidney disease (UAKD). Although uromodulin accumulates in the kidney of UAKD patients, it is unclear whether this is the wildtype or mutant form. In this study, we established a liquid chromatography (LC)-mass spectrometry/mass spectrometry (MS/MS)-based method for the detection of uromodulin mutants, using the C148W mutant as a target molecule. Membrane and cytosolic fractions of kidney samples from transgenic (Tg) mice expressing the C148W uromodulin mutant were shown to contain human uromodulin by western blotting, and mutant uromodulin with the C148W mutant sequence was observed by proteomic and selected reaction monitoring analyses. Our LC-MS/MS-based method is therefore useful for detection of mutant uromodulin that is undetectable by western blotting alone.
authors: Department of Analytical Chemistry, School of Pharmaceutical Sciences, Teikyo University, Kanagawa, Japan. myasuda@pharm.teikyo-u.ac.jp
authors: Makoto
authors: M
authors: Yasuda
authors: Kiyoko
authors: K
authors: Kaneko
authors: Kiyoka
authors: K
authors: Kaneko
authors: Hiroka
authors: H
authors: Hachisu
authors: Noriko
authors: N
authors: Yamaoka
authors: Ken-ichi
authors: K
authors: Mawatari
authors: Kazuya
authors: K
authors: Nakagomi
authors: Norihiko
authors: N
authors: Minoura
authors: Makoto
authors: M
authors: Hosoyamada
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007668:Kidney / Q000378:metabolism; D008297:Male; D013058:Mass Spectrometry; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D009154:Mutation / Q000235:genetics*; D058949:Uromodulin / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D050505:Mutant Proteins; D058949:Uromodulin