ELEMENTARY STEPS OF CHAPERONIN PROMOTED PROTEIN FOLDING

描述：这是一个在第4-8年内继续支持的应用程序 旨在定义分子的作用机制的研究 伴侣GroEL可以帮助变性蛋白质的折叠。所有的 这项研究是为了阐明单个周期的基本步骤 伴侣促进了蛋白质的折叠。许多不同的实验 提出了从定点突变到X射线的各种技术 结晶学和中子散射。表面等离子激元的应用 共振在这些实验中得到了很好的应用。拟议的研究 将测试3个具体目标：1)评估GroEL模型的预测 以核苷酸为基础的功能促进多肽链的释放。 例如，使用GroEL的单环变体将测试 预测与多肽结合的环与核苷酸结合较弱， 但促进非原生链的快速释放。此外，它将是 已确定单个环的全核苷酸占有率是否为 释放非本机链所需的。方法包括 确定核苷酸促进的流体力学和散射研究 构象变化以及它们是否协调一致并遵循 用核苷酸饱和。

DESCRIPTION: This is an application to continue support in years 4-8 for studies aimed at defining the mechanisms by which the molecular chaperone, GroEL, can assist the folding of denatured proteins. All of the research is to elucidate elementary steps of a single cycle of chaperonin facilitated protein folding. Many different experimental techniques are proposed ranging from site directed mutagenesis to X-ray crystallography and neutron scattering. The use of surface plasmon resonance is well used in these experiments. The proposed research would test 3 specific aims: 1) evaluate predictions of a model for GroEL function based on nucleotide promoted release of polypeptide chains. For example, the use of single-ring variants of GroEL would test the prediction that the ring with bound polypeptide binds nucleotides weakly, but promotes rapid non-native chain release. Further, it will be determined whether full nucleotide occupancy of a single ring is required for release of non-native chains. Methods would include hydrodynamic and scattering studies to determine nucleotide promoted conformational changes and whether they are concerted and follow saturation with nucleotide.