| Title: | Proteomic identification of a large complement of rat urinary proteins |
| Author(s): | Thongboonkerd V; Klein JB; Arthur JM; |
| Address: | "Core Proteomics Laboratory, Kidney Disease Program, Department of Medicine, University of Louisville, Louisville, Ky 40202, USA. thongboonkerd@dr.com" |
| ISSN/ISBN: | 1660-2129 (Electronic) 1660-2129 (Linking) |
| Abstract: | "The characterization of urinary proteins is an important tool to identify disease-related biomarkers and to better understand renal physiology. Expression of urinary proteins has been previously studied by Western blotting and other immunological methods. The scope of such studies, however, is limited to previously identified proteins for which specific antibodies are existed. We used proteomic analysis to identify proteins and to construct a proteome map for Sprague-Dawley (SD) rat urine isolated by ultracentrifugation. Urinary proteins were separated by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and visualized by silver staining. Proteins were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), followed by peptide mass fingerprinting using the NCBI protein database. A total of 350 protein spots were visualized. From 250 excised spots, 111 protein components were identified including transporters, transport regulators, chaperones, enzymes, signaling proteins, cytoskeletal proteins, pheromone-binding proteins, receptors, and novel gene products. The presence of a number of these identified proteins was unexpected and had not previously been identified in the urine. 2-D Western blot analyses for randomly selected proteins (ezrin, HSP70, beta- and gamma-actin, Rho-GDI, and l-myc) clearly confirmed the proteomic identification. Several potential posttranslational modifications were predicted by bioinformatic analyses. These data indicate that a large complement of expected and unexpected urinary proteins can be simultaneously studied by proteomic analysis. This approach may lead to better understanding of renal physiology and pathophysiology, and to biomarker discovery" |
| Keywords: | "Actins/urine Amino Acid Sequence Animals Electrophoresis, Gel, Two-Dimensional Male Molecular Sequence Data Peptide Mapping Proteinuria/*urine *Proteomics/methods Rats Rats, Sprague-Dawley Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Ur;" |
| Notes: | "MedlineThongboonkerd, Visith Klein, Jon B Arthur, John M eng R01 HL66358-01/HL/NHLBI NIH HHS/ R21 DK62086-01/DK/NIDDK NIH HHS/ Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. Switzerland 2003/11/12 Nephron Exp Nephrol. 2003; 95(2):e69-78. doi: 10.1159/000073674" |
