title: Development of LC-MS method for detection of mutant uromodulin protein.
abstract: Mutations in the uromodulin gene cause the autosomal disorders familial juvenile hyperuricemic nephropathy (FJHN) and medullary cystic kidney disease type 2 (MCKD2). However, methods to detect the mutant form of the uromodulin protein have not been developed. In this study, we developed a liquid chromatography-mass spectrometry (LC-MS) method for detection of the mutated uromodulin peptide (C148W). Our method can distinguish the mutant peptide, GWHWE, from wildtype peptide, GWHC*E. Using MS/MS analysis with a selected reaction monitoring (SRM) mode, peptide-specific fragment ions (m/z 714 --> 381, 471, 567, and 679 for GWHWE and m/z 688 --> 381, 445, 541, and 653 for GWHC*E) were detected.
authors.affiliation: Laboratory of Analytical Chemistry, School of Pharmaceutical Sciences, Teikyo University, Kanagawa, Japan. myasuda@pharm.teikyo-u.ac.jp
authors.forename: M
authors.initials: M
authors.lastname: Yasuda
authors.forename: K
authors.initials: K
authors.lastname: Kaneko
authors.forename: H
authors.initials: H
authors.lastname: Hachisu
authors.forename: M
authors.initials: M
authors.lastname: Ochiai
authors.forename: T
authors.initials: T
authors.lastname: Yamanobe
authors.forename: K
authors.initials: K
authors.lastname: Mawatari
authors.forename: K
authors.initials: K
authors.lastname: Nakagomi
authors.forename: N
authors.initials: N
authors.lastname: Minoura
authors.forename: M
authors.initials: M
authors.lastname: Hosoyamada
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods*; D004252:DNA Mutational Analysis / Q000379:methods*; D006801:Humans; D009088:Mucoproteins / Q000235:genetics*; D050505:Mutant Proteins / Q000235:genetics*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D058949:Uromodulin
chemical_list: D009088:Mucoproteins; D050505:Mutant Proteins; C548612:UMOD protein, human; D058949:Uromodulin