BACKGROUND: Western blotting has previously been used to identify changes in protein expression in renal tissue. However, only a few proteins can be studied in each experiment by Western blot. We have used proteomic tools to construct protein maps of rat kidney cortex and medulla. METHODS: Expression of proteins was determined by silver stain after two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Protein spots were excised and digested with trypsin. Peptide masses were identified by MALDI-TOF mass spectrometry. The Mascot search engine was used to analyze the peptide masses and identify the proteins. RESULTS: Seventy-two proteins were identified (54 unique proteins) out of approximately 1000 spots visualized on each gel. Most of the spots were expressed both in cortex and medulla. Of the identified proteins, three were expressed only in medulla and one only in cortex. Nine proteins were expressed in both regions but to a greater extent in cortex and three proteins were expressed more in medulla. Differential expression was confirmed for three proteins by Western blot. CONCLUSIONS: A large group of proteins and their relative expression levels from cortical and medullary portions of rat kidneys were found. Sixteen proteins are differentially expressed. Proteomics can be used to identify differential expression of proteins in the kidney on a large scale. Proteomics should be useful to detect changes in renal protein expression in response to a large range of physiological and pathophysiological stimuli.
BACKGROUND: Western blotting has previously been used to identify changes in protein expression in renal tissue. However, only a few proteins can be studied in each experiment by Western blot. We have used proteomic tools to construct protein maps of rat kidney cortex and medulla. METHODS: Expression of proteins was determined by silver stain after two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Protein spots were excised and digested with trypsin. Peptide masses were identified by MALDI-TOF mass spectrometry. The Mascot search engine was used to analyze the peptide masses and identify the proteins. RESULTS: Seventy-two proteins were identified (54 unique proteins) out of approximately 1000 spots visualized on each gel. Most of the spots were expressed both in cortex and medulla. Of the identified proteins, three were expressed only in medulla and one only in cortex. Nine proteins were expressed in both regions but to a greater extent in cortex and three proteins were expressed more in medulla. Differential expression was confirmed for three proteins by Western blot. CONCLUSIONS: A large group of proteins and their relative expression levels from cortical and medullary portions of rat kidneys were found. Sixteen proteins are differentially expressed. Proteomics can be used to identify differential expression of proteins in the kidney on a large scale. Proteomics should be useful to detect changes in renal protein expression in response to a large range of physiological and pathophysiological stimuli.
Authors: Cheryl Smith; Michael Merchant; Andrea Fekete; Ha-Long Nyugen; Paul Oh; You-Lin Tain; Jon B Klein; Chris Baylis Journal: Nephrol Dial Transplant Date: 2008-12-10 Impact factor: 5.992
Authors: Mehmet M Altintas; Kumiko Moriwaki; Changli Wei; Clemens C Möller; Jan Flesche; Jing Li; Suma Yaddanapudi; Mohd Hafeez Faridi; Markus Gödel; Tobias B Huber; Richard A Preston; Jean X Jiang; Dontscho Kerjaschki; Sanja Sever; Jochen Reiser Journal: J Biol Chem Date: 2014-05-09 Impact factor: 5.157
Authors: Vladimir A Valera; Elsa Li-Ning-T; Beatriz A Walter; David D Roberts; W M Linehan; Maria J Merino Journal: J Cancer Date: 2010-10-14 Impact factor: 4.207