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: 6120533
• 负责人: PAUL B SIGLER
• 金额: $ 0.02万
• 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/6120533
• 关键词: bioimaging /biomedical imaging; 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.

我们的实验室在过去的一年中进行了国际象棋A1，F1 和F2，NSLS X4 A，X12和X25，以及APS，以扩展分辨率 并改善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。 在这些旅行中，我们 获得了大量完整的高分辨率数据集， 直接用于确定下列新结构：（1） 与孕酮复合的孕酮配体结合结构域;（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; 嗜热菌EF-Tu/EF-Ts复合物;和（5）沃氏青霉三元复合物 TATA结合蛋白TFIIB和含有TFIIB的DNA序列， 共识TATA框2.1
(see出版物）。 数据最近 在国际象棋中收集的信息有助于解决上述所有问题 结构. 此外，还发表了一篇关于分子机制的论文， 磷酸化在异三聚体调节中的作用 G-Proteins by Escherducin也在准备从收集的数据， 对磷酸化形式的光磷蛋白进行国际象棋。 的其他项目 仍在进行中，受到 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：了解蛋白质折叠的能力。 ATP驱动GroEL-GroES反应循环，我们希望研究相关的 ATP酶循环中的中间体。 GroEL/GroES/（ADP）7/（AlFx）7 晶体结晶度为4
但只有初步测试 今年的F1国际象棋比赛。 寻找最佳冷冻条件 条件正在进行中。 牛视觉抑制：抑制 晶体膨胀至3.0
同步辐射分辨 X-25光束线和MacCHESS A-1光束线）。 我们已经收集了完整的 原生数据集到3.1
（C2221; a = 169
，B = c = 191
晶体尺寸： 0.2 x 0.2 x 0.03 mm3）。 有用的MAD、重原子和反常数据集 在CHESS、X-25和APS收集了用于定相的数据。 季 VP 16酸性激活结构域与hTBPc、hTFIIB复合的复合物 启动子DNA 迄今为止，没有高分辨率结构信息 存在于结合到 起始前复合体 因此，这种晶体结构将揭示 强酸性激活结构域和 基础机制，导致转录激活。 的 晶体的折射率约为4.0
在一个家庭来源（在100 K）。 几 2.6
原始数据集（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 mm 3） 国际象棋F2和F1线。