STRUCTURE AND DYNAMICS OF PKA

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。所列机构为 中心，不一定是研究者的机构。 远程位点之间的变构通信是酶功能中的关键要素。使用基于TROSY的NMR光谱，我们将绘制长程变构变化沿着的催化亚基的cAMP依赖性蛋白激酶A（PKA）的骨架结合后的核苷酸类似物（AMP-PNP）和肽LRRASLG（Kemptide）作为底物。 将获得两个配体的结合常数以及开和关速率的计算。 由于酶保留活性在我们的NMR条件下，在这项研究中监测的结构变化将反映在营业额的构象的相互转换。 这些研究代表了多结构域激酶营业额如何涉及其结构内传输的复杂动态的一个范例。此外，这是第一次报告的监测动态变化的活性激酶在原子分辨率。

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.