ELECTROSTATIC STABILIZATION OF A PROTEIN-DNA COMPLEX

蛋白质-DNA 复合物的静电稳定性

基本信息

批准号：
6525815
负责人：
SHARI J SPECTOR
金额：
$ 3.03万
依托单位：
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-08-01 至 2003-09-12
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6525815
关键词：
DNA binding protein DNA replication chemical stability chromatin circular dichroism gel mobility shift assay gene complementation gene expression gene mutation intermolecular interaction ionic bond mutant nucleic acid sequence protein denaturation protein purification protein structure function site directed mutagenesis transcription factor

项目摘要

Protein-DNA complexes are extremely important in biology, controlling chromatin structure, DNA replication, and protein expression. In fact, mutations in transcriptional activators and repressors or in proteins involved in DNA replication are common causes of cancer. A greater understanding of the interactions that stabilize these complexes will improve our understanding of the disease states that result from mutations to DNA binding proteins. The goal of the project described in this proposal is to identify a set of general rules describing the role of electrostatics in the stability and binding affinity of protein- DNA complexes. A combination of theoretical and experimental approaches will be used to meet this goal. Continuum electrostatic calculations can suggest mutations that will improve the electrostatic complementarity in the Arc repressor- DNA complex, and the predicted mutations will be tested experimentally. In addition, mutations that improve Arc repressor-DNA binding will also be identified by combinatorial mutagenesis techniques, and calculations will be performed to analyze the changes in the electrostatic interactions in the complex.

蛋白质-DNA复合物在生物学，控制染色质结构，DNA复制和蛋白质表达方面非常重要。实际上，转录激活因子和阻遏物或参与DNA复制的蛋白质的突变是癌症的常见原因。对稳定这些复合物的相互作用的更深入的了解将改善我们对突变与DNA结合蛋白产生的疾病状态的理解。本提案中描述的项目的目的是确定一组描述静电在蛋白质DNA复合物稳定性和结合亲和力中的作用的通用规则。理论方法和实验方法的结合将用于实现这一目标。连续静电计算可以表明突变可以改善电弧抑制剂DNA复合物中的静电互补性，并且预测突变将进行实验测试。此外，还将通过组合诱变技术来鉴定改善ARC抑制剂-DNA结合的突变，并将进行计算以分析复合物中静电相互作用的变化。