An Innovative Approach for Profiling Protein Kinase Substrates

描述(由申请人提供)： 信号转导是几乎所有生物过程的中心组成部分。蛋白激酶通过磷酸化参与信号放大的蛋白以及执行细胞对外部或内部刺激的反应，在信号通路中发挥关键作用。异常的激酶信号在几乎所有癌症和数百种其他疾病的病因中扮演着重要的角色。用系统的方法阐明由激酶驱动的信号通路有可能阐明新的治疗干预点。生理激酶底物的鉴定是这一努力的重要组成部分。目前，需要能够广泛部署的技术来解决这一问题。我们正在开发一种简单、直接的激酶底物分析技术，我们称之为反向凝胶内激酶分析。这种分析方法能够在整个或预分选的复杂生物提取物中发现激酶底物。如果开发和验证得当，这种分析方法有可能通过多个实验室的共同努力，能够快速识别许多酶的潜在生理底物。

DESCRIPTION (provided by applicant): Signal transduction is a central component of nearly all biological processes. Protein kinases play critical roles in signaling pathways by phosphorylating proteins involved in signal amplification and in executing the cellular response to extrinsic or intrinsic stimuli. Aberrant kinase signaling plays a role in the etiology of nearly all cancers and in hundreds of other diseases. Systematic approaches to the elucidation of the signaling pathways driven by kinases have the potential to illuminate new points of therapeutic intervention. The identification of physiologic kinase substrates is an important component of this endeavor. Currently, there is a need for technologies that can be widely deployed to approach this problem. We are developing a simple, straightforward technique for kinase substrate profiling that we term the reverse in-gel kinase assay. This assay enables the discovery of kinase substrates in whole or pre-fractionated complex biological extracts. If properly developed and validated, this assay has the potential, through the concerted effort of multiple laboratories, to enable rapid identification of potential physiologic substrates of many kinases.