STRUCTURE/FUNCTION OF HIV TAT-TAR COMPLEX

HIV TAT-TAR 复合物的结构/功能

• 批准号: 2673004
• 负责人: TARIQ M RANA
• 金额: $ 25.36万
• 依托单位: UNIV OF MED/DENT NJ-R W JOHNSON MED SCH
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-01 至 2002-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2673004
• 关键词: human immunodeficiency virus 1; intermolecular interaction; protein structure function; structural biology; virus RNA; virus protein

The PI plans to use the TAT-TAR system for development of techniques to probe the 3-dimensional structure of protein-RNA complexes using a variety of novel physical and chemical approaches. He has developed techniques to link chelators to unique nucleotides in chemically synthesized RNA that permit determination of nearby residues by indentification of the sites of cleavage mediated by the metal-chelator conjugate. This technology will also be used to monitor conformational changes that occur upon binding of protein factors. Specific residue contacts in bound proteins iwll be determined by ribonucleotide-linked celators that serve as proteases, through photo-crosslinking and through the use of chelating amino acid analogs inserted into specific sites of TAR binding TAT peptides. In this latter approach contacts will be determined by clelate-mediated cleavage of the bound RNA. The PI will also convert chelators bound to specific RNA sites to fluorescence energy donors for transfer to acceptors attached to Tat peptides. The distances determined by these methods will be used to set constraints on possible RNA-protein structures.

PI计划使用TAT-TAR系统进行技术开发 使用 各种新颖的物理和化学方法。 他已经开发出 将螯合剂与独特核苷酸进行化学连接的技术 合成的RNA允许通过以下方式测定附近的残基 金属螯合剂介导的裂解位点的鉴定 共轭。 该技术还将用于监测构象 蛋白质因子结合后发生的变化。 特定残留量 结合蛋白中的接触将通过核糖核苷酸连接来确定 通过光交联充当蛋白酶的celators 通过使用插入特定的螯合氨基酸类似物 TAR 结合 TAT 肽的位点。 在后一种方法中，接触 将通过细胞介导的结合RNA的切割来确定。 这 PI 还将结合特定 RNA 位点的螯合剂转化为 荧光能量供体转移至连接到 Tat 的受体 肽。 通过这些方法确定的距离将用于 对可能的 RNA-蛋白质结构设置限制。