{"id":55,"date":"2006-12-14T12:38:52","date_gmt":"2006-12-14T16:38:52","guid":{"rendered":"https:\/\/www.bumc.bu.edu\/cardiovascularproteomics\/project-4\/"},"modified":"2019-07-01T13:50:22","modified_gmt":"2019-07-01T17:50:22","slug":"project-4","status":"publish","type":"page","link":"https:\/\/www.bumc.bu.edu\/cardiovascularproteomics\/project-4\/","title":{"rendered":"Project 4"},"content":{"rendered":"

Post-translational Modification of Vascular Proteins by Homocysteine<\/h3>\n

We previously demonstrated that elevated homocysteine evokes vascular oxidant stress, both by generating reactive oxygen species via thiol oxidation as well as suppressing expression of glutathione peroxidase-1 (GPx-1) in vascular cells. This project will expand these observations to determine the specific protein targets and the specific oxidative modifications that occur in the setting of hyperhomocysteinemia. Our aims are:<\/p>\n

    \n
  1. Use cultured human fibroblasts deficient in cystathionine-beta-synthase, and incubate them with increasing concentrations of methionine, hypoxanthine, and aminopterin (to inhibit remethylation) to obtain graded increases in intracellular homocysteine. Using the ICAT reagents to be developed and other proteomic techniques, proteins will be identified with oxidation of thiols, carbonylation, 3-nitrotyrosine formation, S<\/em>-nitrosation, and homocysteinylation.<\/li>\n
  2. Examine the proteome in cystathionine-beta-synthase knockout mice.<\/li>\n
  3. Devise and test an ICAT reagent specific for homocysteinylated proteins.<\/li>\n
  4. Apply reverse engineering schemes to quantify the effects of homocysteine on vascular oxidant stress in the fibroblasts mentioned above, and then in the knockout mouse model. This will allow us to predict the effects of changing the oxidation states of protein thiols on the rest of the proteome and on cell function. We expect that this will confirm known functional effects of thiol oxidation and to uncover new effects that may then be tested experimentally. Using the results obtained, we expect to be able to understand other cell and mouse models of oxidant stress, as well as to apply the results to proteomic analyses performed in patients.<\/li>\n<\/ol>\n

    \"schematic<\/p>\n

    Homocysteine Metabolism<\/a> \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \"\"Interim update 2006<\/a>\u00a0 \u00a0 Interim update 2005<\/a><\/p>\n

     <\/p>\n

    \"imageDiane E. Handy, Ph.D.<\/a>, Assistant Professor of Medicine, Harvard Medical School and Associate Biochemist, Brigham and Women’s Hospital<\/p>\n

     <\/p>\n

     <\/p>\n

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    \"image<\/a><\/p>\n

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    Joseph Loscalzo, M.D., Ph.D.<\/a>, Hersey Professor of the Theory and Practice of Medicine, Harvard Medical School and Chairman, Department of Medicine, Brigham and Women’s Hospital.<\/p>\n","protected":false},"excerpt":{"rendered":"

    Post-translational Modification of Vascular Proteins by Homocysteine We previously demonstrated that elevated homocysteine evokes vascular oxidant stress, both by generating reactive oxygen species via thiol oxidation as well as suppressing expression of glutathione peroxidase-1 (GPx-1) in vascular cells. This project will expand these observations to determine the specific protein targets and the specific oxidative modifications […]<\/p>\n","protected":false},"author":2,"featured_media":0,"parent":0,"menu_order":74,"comment_status":"closed","ping_status":"closed","template":"","meta":[],"_links":{"self":[{"href":"https:\/\/www.bumc.bu.edu\/cardiovascularproteomics\/wp-json\/wp\/v2\/pages\/55"}],"collection":[{"href":"https:\/\/www.bumc.bu.edu\/cardiovascularproteomics\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/www.bumc.bu.edu\/cardiovascularproteomics\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/www.bumc.bu.edu\/cardiovascularproteomics\/wp-json\/wp\/v2\/users\/2"}],"replies":[{"embeddable":true,"href":"https:\/\/www.bumc.bu.edu\/cardiovascularproteomics\/wp-json\/wp\/v2\/comments?post=55"}],"version-history":[{"count":9,"href":"https:\/\/www.bumc.bu.edu\/cardiovascularproteomics\/wp-json\/wp\/v2\/pages\/55\/revisions"}],"predecessor-version":[{"id":2679,"href":"https:\/\/www.bumc.bu.edu\/cardiovascularproteomics\/wp-json\/wp\/v2\/pages\/55\/revisions\/2679"}],"wp:attachment":[{"href":"https:\/\/www.bumc.bu.edu\/cardiovascularproteomics\/wp-json\/wp\/v2\/media?parent=55"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}