SPECIFIC PROTEIN DNA COMPLEXES TO STUDY CHEMICAL MECHANISM IN CELLULAR REG

用于研究细胞调节化学机制的特定蛋白质 DNA 复合物

• 批准号: 6281306
• 负责人: PAUL B SIGLER
• 金额: $ 9.21万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-15 至 1999-08-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6281306
• 关键词: biological products; biomedical resource; proteins; radiography; structural biology

Our laboratory has in the past year conducted runs at CHESS A1, F1 and F2, NSLS X4A, X12 and X25, as well as APS to extend the resolution and improve the quality of diffraction data for crystalline complexes involved in chaperonin mediated protein folding, signal transduction, transcriptional and translational regulation. During those trips we have obtained numerous complete high resolution data sets resulting directly in the determination of the following new structures: (1) progesterone ligand binding domain complexed with progesterone; (2) human estrogen receptor ligand binding domain complexed with estradiol; (3) the chaperonin GroEL/GroES/ADP complex; (4) the T. thermophilus EF-Tu/EF-Ts complex; and (5) a P. woesei ternary complex of the TATA-binding protein, TFIIB, and a DNA sequence containing a consensus TATA box at 2.1
(see publications section). Data recently collected at CHESS has been instrumental in solving all of the above structures. In addition a paper on the molecular mechanism for the role of phosphorylation in the regulation of hetereotrimeric G-Proteins by phosducin is also in preparation from data collected at CHESS on the phosphorylated form of phosducin. Other projects which are still in progress which have been significantly influenced by CHESS synchrotron data collection include the following: Chaperonin-Medicated Protein Folding: To understand the capacity of ATP to drive the GroEL-GroES reaction cycle, we wish to study relevant intermediates in the ATPase cycle. GroEL/GroES/(ADP)7/(AlFx)7 crystals diffract to 4
at our home source but only initial tests were done at the CHESS F1 line this year. Searching for optimal freezing conditions is in progress. Bovine Visual Arrestin: The arrestin crystals diffract to 3.0
resolution using synchrotron radiation (NSLS X-25 beamline and MacCHESS A-1 beamline). We have collected complete native data sets to 3.1
(C2221; a = 169
, b = c = 191
; crystal size: 0.2 x 0.2 x 0.03 mm3). Useful MAD, heavy atom and anomalous data sets for phasing have been collected at CHESS, X-25 and APS. Quaternary Complex of VP16 Acidic Activation Domain Complexed with hTBPc, hTFIIB on Promoter DNA. To date no high resolution structural information exists on a transcriptional activator domain bound to the preinitiation complex. Hence, this crystal structure will reveal the essential contacts made between a strong acidic activation domain and the basal machinery, resulting in transcriptional activation. The crystals diffract to about 4.0
on a home source (at 100 K). Several 2.6
native data sets (P21; a = 118.7
, b = 122.2
, c = 140.8
, b = 113.0 ; crystal size: 0.4 x 0.1 x 0.05 mm3) were collected recently at CHESS F2 and F1 lines.

我们的实验室去年在 CHESS A1、F1 上进行了运行 和 F2、NSLS X4A、X12 和 X25，以及 APS 来扩展分辨率 并提高晶体复合物的衍射数据的质量 参与伴侣蛋白介导的蛋白质折叠、信号转导、 转录和翻译调控。 在那些旅行中我们 已获得大量完整的高分辨率数据集 直接用于确定以下新结构： (1) 与黄体酮复合的黄体酮配体结合域； (2) 人雌激素受体配体结合结构域与 雌二醇； (3) 伴侣蛋白 GroEL/GroES/ADP 复合物； （4）T。 嗜热菌 EF-Tu/EF-Ts 复合物； (5) P. woesei 三元复合体 TATA 结合蛋白 TFIIB 和含有 TATA 共识框为 2.1（参见出版物部分）。 最近数据 CHESS 收集的信息有助于解决上述所有问题 结构。 另外一篇关于分子机制的论文 磷酸化在异源三聚体调节中的作用 G-Proteins by phosducin 也正在根据收集的数据进行制备 CHESS 对磷酸化形式的磷酸化蛋白的影响。 其他项目其中 仍在进行中，并受到显着影响 CHESS同步加速器数据采集包括以下内容： 伴侣蛋白介导的蛋白质折叠：了解 ATP驱动GroEL-GroES反应循环，我们希望研究相关 ATP酶循环的中间体。 GroEL/GroES/(ADP)7/(AlFx)7 在我们的家乡，晶体衍射为 4，但仅进行了初步测试 今年在 CHESS F1 线上完成。 寻找最佳冷冻方法 条件正在进行中。 牛视觉抑制素：抑制素 使用同步加速器辐射 (NSLS X-25 光束线和 MacCHESS A-1 光束线）。 我们收集了完整的 原始数据集为 3.1（C2221；a = 169，b = c = 191；晶体尺寸： 0.2×0.2×0.03 毫米3）。 有用的 MAD、重原子和异常数据集 已在 CHESS、X-25 和 APS 收集了用于分相的信息。 第四纪 VP16 酸性激活结构域与 hTBPc、hTFIIB 的复合物 在启动子 DNA 上。 迄今为止还没有高分辨率的结构信息 存在于与 预引发复合体。 因此，该晶体结构将揭示 强酸性激活结构域和 基础机制，导致转录激活。 这 晶体在家用光源（100 K）上的衍射约为 4.0。 一些 2.6 原生数据集 (P21; a = 118.7 , b = 122.2 , c = 140.8 , b = 113.0;晶体尺寸：0.4 x 0.1 x 0.05 mm3) 最近收集 在 CHESS F2 和 F1 线。