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Detection of protein Z in a renal calculus composed of calcium oxalate monohydrate with the use of liquid chromatography-mass spectrometry/mass spectrometry following two-dimensional polyacrylamide gel electrophoresis separation
Authors:Kaneko Kiyoko  Yamanobe Tomoyo  Nakagomi Kazuya  Mawatari Ken-ichi  Onoda Maki  Fujimori Shin
Institution:Central Laboratory of Analytical Biochemistry, Teikyo University, Tokyo 173-8605, Japan. kikaneko@pharm.teikyo-u.ac.jp
Abstract:Protein Z, a vitamin K-dependent plasma protein, has been detected for the first time in a renal calculus along with osteopontin and prothrombin. The renal calculus was obtained from a hyperuricemic patient. Following two-dimensional polyacrylamide gel electrophoresis, the calculus was analyzed with the use of liquid chromatography mass spectrometry (LC-MS). The spectrometer was equipped with a nanoelectrospray interface and an ion trap. Four peptides were determined from a protein in the calculus through LC-MS/MS analysis. Tandem mass spectrum database matching tools were used to identify the protein as protein Z. Authentic protein Z was also analyzed using the same method, and all four peptides determined in the calculus were similarly identified. Whereas protein Z has been reported to be one of the vitamin K-dependent calcium-binding proteins, its role has not been well established. The fact that protein Z exists in a renal calculus composed of calcium oxalate will be beneficial in any future investigations into its role in the body.
Keywords:Protein Z  Renal calculus  2D-PAGE separation  LC-MS/MS  Micro-area X-ray diffractometer  Calcium oxalate monohydrate
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