STRUCTURE AND DYNAMICS OF PKA

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 远端位点之间的变构通讯是酶功能中的一个关键因素。使用基于TROSY的核磁共振波谱，我们将绘制cAMP依赖的蛋白激酶A(PKA)催化亚单位(PKA)与核苷酸类似物(AMP-PNP)和多肽LRRASLG(Kemptie)结合时沿其主干的长程变构变化。将获得两个配体的结合常数以及开启和关闭速率的计算。由于酶在我们的核磁共振条件下保持活性，本研究中监测的结构变化将反映周转过程中构象的相互转换。这些研究代表了多结构域激酶周转如何涉及在其结构内传递的复杂动力学的范例。此外，这是第一个在原子分辨率下监测到活性激酶内动态变化的报告。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Allosteric communication between remote sites is a crucial element in enzymatic function. Using TROSY-based NMR spectroscopy, we will map long-range allosteric changes along the backbone of the catalytic subunit of cAMP-dependent protein kinase A (PKA) upon binding a nucleotide analogue (AMP-PNP) and the peptide LRRASLG (Kemptide) as substrates. Calculations for the binding constants of the two ligands, as well as for on and off rates will be obtained. Since the enzyme retains activity under our NMR conditions, the structural changes monitored in this study will reflect the interconversion of conformations during turnover. These studies represent a paradigm for how multidomain kinase turnover involves complex dynamics transmitted within its structure. In addition, this is the first report of monitored dynamic changes within an active kinase at atomic resolution.