SPECTROSCOPY OF HEAVY ATOM PERTURBED BIOPOLYMERS

重原子扰动生物聚合物的光谱

• 批准号: 6055890
• 负责人: AUGUST H MAKI
• 金额: $ 16.99万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-01-01 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6055890
• 关键词: DNA binding protein; RNA; electron spin resonance spectroscopy; genetic operator element; heavy metals; human immunodeficiency virus 1; intermolecular interaction; nucleic acid sequence; nucleocapsid; nucleoproteins; phosphorescence; protein structure function; reporter genes; simian immunodeficiency virus; structural biology; triplet state; virus protein

Protein-nucleic acid interactions play essential roles in cellular processes and in the functioning of mammalian retroviruses. To study these interactions the triplet states of aromatic residues will be studied using phosphorescence and optical detection of magnetic resonance (ODMR). Alloproteins will incorporate Trp, Phe, and Tyr analogs that can be selectively excited at the red edge in nucleic acid complexes. Of importance is uncovering factors that influence the stability and sequence selectivity of nucleic acid binding and the extent to which they are influenced by aromatic stacking. The primary focus will be HIV-1 and SIV. Foremost will be studies of the interactions of nucleocapsid (NC) proteins with the RNA genome during maturation in RNA encapsidation. The NC proteins are characterized by conserved Cys-X2-Cys-X4-His-X4-Cys retroviral zinc finger motifs containing aromatic residues at specific sites. Zinc fingers have been found to undergo base-stacking with model RNAs. Similar interactions will be sought. RNA substrates will be extended to sequences containing psi, the encapsidation region of genomic RNA. Spin-lattice relaxation of the triplet state will be studied as a possible reporter of structure. Effects of inducer and operator binding on single-W constructs of E. coli lac repressor will be studied. ODMR or interleukin-1-beta alloproteins will be used to test effects of Trp analog incorporation on protein structure.

蛋白质-核酸相互作用在细胞中发挥重要作用 哺乳动物逆转录病毒的过程和功能。 学习 这些相互作用芳香残基的三重态将是 研究人员利用磷光和光学检测磁性 共振（ODMR）。 同种蛋白将包含 Trp、Phe 和 Tyr 可以在核酸红边选择性激发的类似物 复合物。重要的是揭示影响因素 核酸结合的稳定性和序列选择性 它们受芳香堆积影响的程度。 初级 重点是 HIV-1 和 SIV。 首要的将是研究 核衣壳 (NC) 蛋白与 RNA 基因组的相互作用 RNA 衣壳化的成熟。 NC 蛋白的特点是 保守的 Cys-X2-Cys-X4-His-X4-Cys 逆转录病毒锌指基序 在特定位点含有芳香族残基。 锌指已 发现与模型 RNA 进行碱基堆积。 类似的互动 将被寻求。 RNA底物将延伸至包含以下的序列 psi，基因组RNA的衣壳化区域。 自旋晶格弛豫 三重态的可能报告者将被研究 结构。 诱导子和操纵子结合对单 W 的影响 将研究大肠杆菌 lac 阻遏物的构建体。 ODMR 或 IL-1-β同种蛋白将用于测试色氨酸的效果 蛋白质结构的类似物掺入。