Chemical approaches for functionally probing the proteome
Chemical approaches for functionally probing the proteome
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DOI：
10.1074/mcp.t100003-mcp200
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发表时间：
2002-01-01
影响因子：
7
通讯作者：
Bogyo, M
中科院分区：
文献类型：
--
作者：
Greenbaum, D;Baruch, A;Bogyo, M
随着全基因组序列的获得，重点已经转向对蛋白质功能的理解。我们开发了一种功能蛋白质组学方法，利用化学反应的荧光探针，通过酶的催化活性来分析和鉴定复杂混合物中的酶。这种方法允许使用单一的二维分离来比较多种酶在各种条件下的活性变化。这些探针还可以用来利用荧光显微镜定位完整细胞中的活性酶。此外，这些探针能够筛选粗裂物或完整细胞中每个酶家族成员的选择性小分子抑制剂。最终，这项技术可以快速识别潜在的药物目标和目标小分子先导化合物。
With the availability of complete genome sequences, emphasis has shifted toward the understanding of protein function. We have developed a functional proteomic methodology that makes use of chemically reactive fluorescent probes to profile and identify enzymes in complex mixtures by virtue of their catalytic activity. This methodology allows a comparison of changes in activity of multiple enzymes under a variety of conditions using a single two-dimensional separation. The probes can also be used to localize active enzymes in intact cells using fluorescence microscopy. Furthermore, the probes enable screens for selective small molecule inhibitors of each enzyme family member within crude lysates; or intact cells. Ultimately, this technology allows the rapid identification of potential drug targets and small molecule lead compounds targeted to them.