MODULATING THE REDUCTIVE UNFOLDING PATHWAY OF RNASE A

调节 RNA酶 A 的还原性解折叠途径

• 批准号: 7721213
• 负责人: HAROLD A. SCHERAGA
• 金额: $ 0.04万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-15 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7721213
• 关键词: Behavior; Bos taurus; Cattle; Computer Retrieval of Information on Scientific Projects Database; Deposition; Endoribonucleases; Funding; Grant; Homologous Gene; Institution; Localized; Location; Modeling; Mutate; Onconase; Pancreatic ribonuclease; Pathway interactions; Predisposition; Protein Dynamics; Proteins; Publishing; Rate; Research; Research Personnel; Resources; Ribonucleases; Site; Source; Structure; United States National Institutes of Health; Variant; Work; design; disulfide bond; molecular dynamics; mutant; protein structure function

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. Reductive unfolding studies of proteins are designed to provide information about intramol. interactions that govern the formation (and stabilization) of the native state and about folding/unfolding pathways. By mutating Tyr92 to G, A, or L in the model protein, bovine pancreatic RNase A, and through analysis of temp. factors and molecular dynamics simulations of the crystal structures of these mutants, it is demonstrated that the markedly different reductive unfolding rates and pathways of RNase A and its structural homolog onconase can be attributed to a single, localized, ring-stacking interaction between Tyr92 and Pro93 in the bovine variant. The fortuitous location of this specific stabilizing interaction in a disulfide-bond-contg. loop region of RNase A results in the localized modulation of protein dynamics that, in turn, enhances the susceptibility of the disulfide bond to redn. leading to an alteration in the reductive unfolding behavior of the homologues. These results have important implications for folding studies involving topol. determinants to obtain folding/unfolding rates and pathways, for protein structure-function prediction through fold recognition, and for predicting proteolytic cleavage sites. Three structures were deposited in the PDB (1YMR, 1YMW, and 1YMN) and the work was published in JACS.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 蛋白质的还原性展开研究旨在提供有关 Intramol 的信息。控制天然状态形成（和稳定）以及折叠/展开途径的相互作用。通过将模型蛋白牛胰腺 RNase A 中的 Tyr92 突变为 G、A 或 L，并通过温度分析。通过对这些突变体晶体结构的因素和分子动力学模拟，证明 RNase A 及其结构同源物 onconase 显着不同的还原解折叠速率和途径可归因于 Tyr92 和 Pro93 之间的单一、局部、环堆积相互作用在牛的变种中。这种特定的稳定相互作用在二硫键结构中的偶然位置。 RNase A 的环区域导致蛋白质动力学的局部调节，进而增强二硫键对 redn 的敏感性。导致同系物还原展开行为的改变。这些结果对于涉及白杨的折叠研究具有重要意义。获得折叠/解折叠速率和途径的决定因素，通过折叠识别进行蛋白质结构功能预测，以及预测蛋白水解切割位点。 三个结构被存放在 PDB（1YMR、1YMW 和 1YMN）中，并且该工作发表在 JACS 上。