Seeking the Biophysical Principles that Govern RTK Activation

项目摘要 受体酪氨酸激酶（RTK）是控制细胞生长的单程跨膜蛋白， 分化、运动和代谢。它们是非常有前途的药物靶点，但对它们的不完全了解 RTKs在细胞膜中的激活机制的不清楚阻碍了有效和安全治疗的发展。 该提案致力于开发新的方法，可以量化分子相互作用， 可以推动这个领域向前发展。在目标1中，我们将开发一个组合的数字和亮度（N&B）和福斯特 共振能量转移（FRET）方法，其产生寡聚体尺寸和解离常数 用于活细胞中的全长RTK。在目标2中，我们将开发一种基于荧光的方法， 活细胞中全长RTK的分子配体结合常数。沿着的是， 通过广泛的科学界的知识，目标中的工作将产生对关键方面的新见解 方法开发中使用的RTK信号。

PROJECT SUMMARY Receptor Tyrosine Kinases (RTKs) are single-pass transmembrane proteins that control cell growth, differentiation, motility, and metabolism. They are very promising drug targets, but the incomplete understanding of the activation mechanism of the RTKs in the membrane hinders the development of effective and safe therapies. This proposal is dedicated to the development of new methodologies that can quantify molecular interactions and can move the field forward. In Aim 1, we will develop a combined Number and Brightness (N&B) and Förster Resonance Energy Transfer (FRET) methodology that yields both the oligomer size and the dissociation constants for full length RTKs in live cells. In Aim 2, we will develop a fluorescence-based methodology that yields molecular ligand binding constants for full-length RTKs in live cells. Along with enabling the acquisition of new knowledge by the broad scientific community, the work in the aims will yield new insights into critical aspects of the signaling by the RTKs used in method development.