STUDIES OF PROTEIN-PROTEIN INTERACTIONS IN NUCLEAR PORE COMPLEXES

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 我们有一个长期的研究计划，以表征一些酶和蛋白质-蛋白质相互作用的动力学。 这包括蛋白质工程方法的发展，包括NMR的片段标记，从NMR弛豫直接确定溶液中的结构域间取向， 在蛋白质-蛋白质结合的细胞研究中，直接和完整地检测蛋白质-蛋白质相互作用表面。 ESR技术通常需要通过提供比NMR方法可用的距离（更大）和时间尺度（更短）来增加用于建模结构和动力学的实验输入。 蛋白酪氨酸激酶（PTKs）：我们专注于结构/动力学的两个方面  (a)活化环磷酸化在PTK活性增强中的作用，和（B）通过活性和定位的分子内/结构域间和分子间变构调节的蛋白-蛋白相互作用的作用。 非受体Csk、Src、Abl和受体PTK FGFR-2目前以多种工程化形式表达，并正在与天然的、常见的疾病相关的多种形式进行研究。 相关的和其它突变/同种型/截短。 我们正在将这些研究扩展到掺入真核生物中，并且ESR潜在地提供了足够的灵敏度来克服是否需要过量的蛋白质掺入来观察NUP/karopherin亲和力的货物修饰的问题。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. We have a long-term program of research to characterize dynamics underlying a number of enzymatic and protein-protein interactions. This has included the development of protein engineering methods including segmental labeling for NMR, the direct determination of interdomain orientations in solution from NMR relaxation, in cell studies of protein-protein binding, and direct, and complete detection of protein-protein interaction surfaces. ESR techniques are generally required to augment the experimental input for modeling structures and dynamics by providing distances (larger) and time scales (shorter) than available from NMR methods. Protein tyrosine kinases(PTKs): We are concentrating on two aspects of structure/dynamics  (a) the role(s) of activation loop phosphorylation in enhanced activity of PTKs, and (b) the role(s) of protein-protein interactions via intramolecular/ interdomain and intermolecular allosteric modulation of activities and localization. The non-receptor Csk, Src, Abl , and receptor PTK FGFR-2 are currently expressed and being studied in multiple engineered forms with both natural, frequently disease related, and other mutations/isoforms/truncations. We are extending these studies to incorporation into eukaryotes, and ESR potentially provides a sufficient sensitivity to overcome the issue of whether excessive protein incorporation is needed to see the cargo modification of NUP/karopherin affinities.