1. Current Proteome Profiling Methods and Applications Yetrib Hathout, Ph.D. Center for Genetic Medicine Children’s National Medical Center & The George Washington University yhathout@cnmcresearch.org

2. The goal on many proteomic studies is determining protein ratios between samples representing different biological states. diseased tissue  healthy tissue drug treated  non-tread drug resistant  non-resistant

3. Reudi Aebersold and Matthias Mann. Nature 2003, 422:198-207 Mass spectrometry Current Methods

4. Database search & protein identification Based peak MS/MS Quantification & Validation Unlabeld Cells sample 1 regular Arg and Lys Labeled Cells sample 2 13 C 6 -Arg & 13 C 6, 15 N 2 -Lys Mix cells1:1 LC-MS and MS/MS analysis Extract proteins And Digest with tryspin Mitochondrial fraction Nuclear fraction ER fraction Zoom scans

5. Applications Understanding cellular and molecular mechanisms of age related macular degeneration. ER stress response and protein trafficking in Vanishing White Matter-Disease. Defining biomarkers and therapeutic targets in pediatric brain tumors.