{"id":59,"date":"2007-06-19T14:37:51","date_gmt":"2007-06-19T18:37:51","guid":{"rendered":"https:\/\/www.bumc.bu.edu\/cardiovascularproteomics\/project-5\/"},"modified":"2012-11-01T12:14:06","modified_gmt":"2012-11-01T16:14:06","slug":"project-5","status":"publish","type":"page","link":"https:\/\/www.bumc.bu.edu\/cardiovascularproteomics\/project-5\/","title":{"rendered":"Project 5"},"content":{"rendered":"

Interim update October 2006<\/a>
\nInterim update 2005<\/a><\/p>\n

Circulating Surrogate Target Cells of Oxidant Stress<\/h3>\n

As a prerequisite for enabling studies of oxidative protein modifications in patients, the purpose of this project is to determine if proteins identified to be oxidatively modified in tissues of the cultured cell and mouse models of oxidant stress are also modified in circulating blood cells. Because the PI has identified oxidative stress in platelets of patients in which cardiovascular risk factors associated with abnormal vascular function, we will study platelets as potential circulating surrogate target cells in which to study oxidative protein modifications. The aims are to:<\/p>\n

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  1. Determine if platelet proteins are oxidatively modified in diabetic LDL receptor knockout mice in which tissue eNOS is oxidized. Platelets from these mice will be examined for NO and superoxide anion release. eNOS will be purified from the platelets, and loss of zinc and oxidation of the zinc thiolate center will be investigated.<\/li>\n
  2. Determine abundance of eNOS and its oxidative modifications in patients with unstable CAD. eNOS protein expression, enzymatic activity, and NO release will be measured. eNOS purified from platelets of these patients will be analyzed for oxidation of the zinc thiolate center, loss of zinc, and other oxidative protein modifications by tryptic digestion and MSn analysis.<\/li>\n
  3. Screen platelet proteins with ICAT reagents developed for oxidative modifications to test their suitability for wider use.<\/li>\n
  4. Screen platelets with known oxidative protein modifications for differences in protein abundance. Bioinformatics analysis of the data will be done to analyze relationships amongst antioxidant and signaling systems.<\/li>\n<\/ol>\n

    \"image<\/p>\n

    Hypothesis:<\/strong> In addition to hemostasis, platelets mediate inflammation and clearance of bacteria from the bloodstream. As with platelet-platelet interactions, platelet-bacteria interactions involve release of granular content and cytoskeletal rearrangements in the platelet. It is not known if the activation of platelets by different pathways leads to differential regulation. The researcher’s working hypothesis is that activation of platelets by thrombotic or immune pathways leads to differential regulation of protein interactions and\/or granule release. To test it, they have chosen FXIIIA as a candidate protein and summarize their work thus far (CPC presentation May 2007 here).<\/a> For FXIIIA-associated proteins<\/strong>:<\/p>\n