Protein Array Technology for Protein Interaction

描述(由申请人提供)：蛋白质结构域通过与相应配体中的短肽基序结合来调节蛋白质之间的相互作用。这些肽识别模块对于组装多蛋白质复合体至关重要。我们已经将聚焦于蛋白质相互作用结构域的GST融合蛋白排列在硝酸纤维涂覆的玻璃片上，以产生蛋白质结构域芯片。本幻灯片上排列的结构域包括WW、SH3、SH2、14-3-3、PDZ、FHA、PH和FF域。我们已经使用多肽证明了排列的结构域保持了它们的结合完整性。此外，我们还表明，蛋白质结构域芯片可以从细胞总裂解液中“钓鱼”蛋白质，然后这些结构域结合的蛋白质可以在芯片上用特定的抗体检测到，从而产生感兴趣的蛋白质的结合图谱。泛经济学公司和德克萨斯大学安德森癌症中心现在计划将这种蛋白质结构域阵列商业化。这种蛋白质芯片不仅将检测蛋白质的数量变化，还有望识别蛋白质配体的质量差异，从而进入信号转导途径的核心。此外，这种类型的蛋白质结构域阵列将有助于在蛋白质水平上识别危险因素和化学预防目标。

DESCRIPTION (provided by applicant): Protein domains mediate protein-protein interaction through binding to short peptide motifs in their corresponding ligands. These peptide recognition modules are critical for the assembly of multi-protein complexes. We have arrayed GST fusion protein, with a focus on protein interaction domains, onto nitrocellulose coated glass slides to generate a protein-domain chip. Domains arrayed on this slide included WW, SH3, SH2, 14-3-3, PDZ, FHA, PH and FF domains. We have demonstrated, using peptides, that the arrayed domains retain their binding integrity. Furthermore, we show that the protein-domain chip can "fish" proteins out of a total cell lysate, these domain bound proteins can then be detected on the chip with a specific antibody, thus producing a binding map for a protein of interest. Panomics, inc. and UT M.D. Anderson Cancer Center now plan to commercialize this protein-domain array. This protein chip will not only detect quantitative changes in proteins, but also promises to identify qualitative differences in protein ligands, thus getting at the heart of signal transduction pathways. In addition, this type of protein-domain array will facilitate the identification of risk factors and chemoprevention targets at the protein level.