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.

我们的实验室在过去的一年中在国际象棋A1进行跑步 和F2，NSLS X4A，X12和X25以及APS以扩展分辨率 并提高晶体复合物的衍射数据质量 参与伴侣蛋白介导的蛋白质折叠，信号转导， 转录和翻译调节。 在那些旅行中 获得了许多完整的高分辨率数据集 直接确定以下新结构：（1） 孕酮配体结合结构域与孕酮复合； （2） 人雌激素受体配体结合结构域与 雌二醇； （3）伴侣蛋白凹槽/凹槽/ADP复合物； （4）T.。 EF-TU/EF-TS复合物； （5）P。Woesei三元络合物 TATA结合蛋白，TFIIB和包含A的DNA序列的 在2.1的共识tata框（请参阅出版物部分）。 最近的数据 在国际象棋中收集的已在解决上述所有方面发挥了作用 结构。 此外，有关分子机制的论文 磷酸化在调节甲甲中三位体中的作用 Phosducin的G蛋白也正在从收集的数据中进行准备 国际象棋磷酸化形式的磷素。 其他项目 仍在进行中 国际象棋同步总数数据收集包括以下内容： 伴侣蛋白药物的蛋白质折叠：了解 ATP驱动凹槽 - 胶质反应周期，我们希望研究相关 在ATPase周期中中间。 groel/groes/（ADP）7/（alfx）7 晶体衍射到我们的家庭源为4，但只有初始测试是 今年在国际象棋F1线上完成。 搜索最佳冻结 条件正在进行中。 牛视觉逮捕：逮捕 使用同步加速器辐射（NSLS）晶体衍射至3.0分辨率 X-25梁线和MacChess A-1梁线）。 我们已经完成了 天然数据集为3.1（C2221; a = 169，b = C = 191;晶体大小： 0.2 x 0.2 x 0.03 mm3）。 有用的疯狂，沉重的原子和异常数据集 为了取代，已在国际象棋，X-25和AP中收集。 第四纪 VP16与HTBPC复合HTFIIB复合的VP16酸性激活结构域 在启动子DNA上。 迄今为止没有高分辨率的结构信息 存在于连接的转录激活器域 预上络合物。 因此，这种晶体结构将揭示 强酸性激活域和 基础机械，导致转录激活。 这 晶体衍射为家庭源（100 K）上约4.0。 一些 2.6本地数据集（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 在国际象棋F2和F1线上。