1. Altered retinoic acid metabolism in diabetic mouse kidney identified by O isotopic labeling and 2D mass spectrometry.
- Author
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Starkey JM, Zhao Y, Sadygov RG, Haidacher SJ, Lejeune WS, Dey N, Luxon BA, Kane MA, Napoli JL, Denner L, and Tilton RG
- Subjects
- Animals, Blotting, Western, Chromatography, High Pressure Liquid methods, Male, Mice, Mice, Inbred C57BL, Microscopy, Fluorescence, Oxygen Isotopes, Reverse Transcriptase Polymerase Chain Reaction, Diabetes Mellitus, Experimental metabolism, Kidney metabolism, Mass Spectrometry methods, Tretinoin metabolism
- Abstract
Background: Numerous metabolic pathways have been implicated in diabetes-induced renal injury, yet few studies have utilized unbiased systems biology approaches for mapping the interconnectivity of diabetes-dysregulated proteins that are involved. We utilized a global, quantitative, differential proteomic approach to identify a novel retinoic acid hub in renal cortical protein networks dysregulated by type 2 diabetes., Methodology/principal Findings: Total proteins were extracted from renal cortex of control and db/db mice at 20 weeks of age (after 12 weeks of hyperglycemia in the diabetic mice). Following trypsinization, (18)O- and (16)O-labeled control and diabetic peptides, respectively, were pooled and separated by two dimensional liquid chromatography (strong cation exchange creating 60 fractions further separated by nano-HPLC), followed by peptide identification and quantification using mass spectrometry. Proteomic analysis identified 53 proteins with fold change >or=1.5 and p
- Published
- 2010
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