Seeking the Biophysical Principles that Govern RTK Activation

项目总结 受体酪氨酸激酶(RTK)是一种控制细胞生长的单程跨膜蛋白， 分化、运动和新陈代谢。他们是非常有希望的药物靶点，但不完全的理解 膜上RTK激活机制的研究阻碍了有效、安全的治疗方法的发展。 这项建议致力于开发新的方法，可以量化分子相互作用和 可以将场向前推进。在目标1中，我们将开发组合数字和亮度(N&B)和Förster 共振能量转移(FRET)方法，同时得出齐聚物大小和解离常数 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，以产生 活细胞中全长RTK的分子配体结合常数。同时支持收购新的 广泛的科学界的知识，AIMS的工作将产生对关键方面的新见解 方法开发中使用的RTK的信令。

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.