Aki Nakayama1, Ryo Kubota2, Minoru Sakatsume3, Hidenori Suzuki4, Akira Katayama5, Kiyoko Kanamori6, Kiyoko Shiba7, Shiro Iijima6,7. 1. Faculty of Health Science Technology, Bunkyo Gakuin University, Tokyo, Japan. anak@bgu.ac.jp. 2. Department of Health Sciences, Saitama Prefectural University, Saitama, Japan. 3. Division of Clinical Nephrology and Rheumatology, Graduate School of Medical and Dental Sciences, Niigata University, Niigata, Japan. 4. Department of Pharmacology, Graduate School of Medicine, Nippon Medical School, Tokyo, Japan. 5. Department of Biochemistry and Molecular Biology, Nippon Medical School, Tokyo, Japan. 6. Faculty of Health Science Technology, Bunkyo Gakuin University, Tokyo, Japan. 7. Division of Health Care Science, Graduate School of Health Care Science, Bunkyo Gakuin University, Tokyo, Japan.
Abstract
BACKGROUND: Analysis of urinary proteins using cellulose acetate membrane electrophoresis (CAME) is a useful and challenging method for the recognition of damaged sites in the kidney. However, protein content of each CAME fraction is still not completely understood. METHODS: In this study, an effective method of protein extraction from each band fractionated by CAME was established, which enabled us to examine the extracted proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and mass spectrometry. RESULTS: Proteins were extracted from the gel and analyzed by mass spectrometry. In all, 31 proteins were identified, including 20 urinary proteins that were newly identified in the CAME-based analysis. CONCLUSION: This methodology was useful for identifying the proteins responsible for creating unique bands on CAME in a urine sample of a patient with drug-induced interstitial nephritis. These findings provide in-depth characterization of urinary protein contents in each CAME fraction.
BACKGROUND: Analysis of urinary proteins using cellulose acetate membrane electrophoresis (CAME) is a useful and challenging method for the recognition of damaged sites in the kidney. However, protein content of each CAME fraction is still not completely understood. METHODS: In this study, an effective method of protein extraction from each band fractionated by CAME was established, which enabled us to examine the extracted proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and mass spectrometry. RESULTS: Proteins were extracted from the gel and analyzed by mass spectrometry. In all, 31 proteins were identified, including 20 urinary proteins that were newly identified in the CAME-based analysis. CONCLUSION: This methodology was useful for identifying the proteins responsible for creating unique bands on CAME in a urine sample of a patient with drug-induced interstitial nephritis. These findings provide in-depth characterization of urinary protein contents in each CAME fraction.
Authors: Michiko Suzuki; Kristina Wiers; Elizabeth B Brooks; Kenneth D Greis; Kathleen Haines; Marisa S Klein-Gitelman; Judyann Olson; Karen Onel; Kathleen M O'Neil; Earl D Silverman; Lori Tucker; Jun Ying; Prasad Devarajan; Hermine I Brunner Journal: Pediatr Res Date: 2009-05 Impact factor: 3.756