STRUCTURAL ANALYSIS OF HUMAN MAIM PROTEIN

人类残害蛋白的结构分析

• 批准号: 8170649
• 负责人: ANN M. STOCK
• 金额: $ 0.49万
• 依托单位: BROOKHAVEN SCIENCE ASSOC-BROOKHAVEN LAB
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8170649
• 关键词: Capital; Computer Retrieval of Information on Scientific Projects Database; Data; Data Set; Escherichia coli; Freezing; Funding; Grant; Home environment; Human; Institution; L-Selenomethionine; Liquid substance; Methods; Muscle; Myosin ATPase; Nitrogen; Optics; Phase; Property; Proteins; Research; Research Personnel; Resolution; Resources; Solvents; Source; Structure; United States National Institutes of Health; indexing; meetings; statistics

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We aim to determine the structure of a human muscle-specific protein containing a Myosin Activation-Interaction Motif (MAIM), a domain with no identifiable sequence similarity to proteins of known structure. The 103 kDa protein has been expressed and purified from E. coli as a soluble active protein. We have generated crystals of both native and SeMet-substituted protein. We have collected complete datasets from native crystals to 2.8 [CAPITAL_A_RING] resolution on our home source (Rigaku MicroMax007 with VariMax HR confocal optics) with good statistics. The data index in a hexagonal spacegroup (a = b = 109.2 [CAPITAL_A_RING], c = 153.7 [CAPITAL_A_RING], [SMALL_ALPHA = [SMALL_BETA] = 90˚, [SMALL_GAMMA] = 120˚) with one molecule in the asymmetric unit and a solvent content of 26%, consistent with the strong diffraction we obtain from relatively small crystals. The SeMet-substituted crystals have diffraction properties analogous to the native crystals, but have been grown to larger size. Crystals have been grown in cryo-ready conditions, frozen, and stored in liquid nitrogen. We wish to collect a complete MAD dataset from SeMet-substituted crystals (22 Met/103 kDa) for solving the phase problem by MAD or SAD methods. Additionally, we will collect a high resolution

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 我们的目的是确定含有肌球蛋白激活相互作用基序（MAIM）的人类肌肉特异性蛋白的结构，该结构域与已知结构的蛋白质无可识别的序列相似。 103 kDa蛋白已从大肠杆菌中表达并纯化为可溶性活性蛋白。 我们已经产生了天然和塞氏取代蛋白的晶体。 我们已经收集了从本地源到2.8 [capital_a_ring]的完整数据集（带有良好统计信息的varimax HR共共聚焦光学器件）上的家用源（rigaku micromax007）。 六边形空间组中的数据索引（a = b = 109.2 [capital_a_ring]，c = 153.7 [capital_a_ring]，[small_alpha = [small_beta] =90˚，[small_gamma] =120˚），在不对称单位中，一个分子在26％的差异中获得一个分子，并获得一个强度的差异。 Semet取代的晶体具有类似于天然晶体的衍射特性，但已生长到更大的尺寸。 晶体已在冷冻的条件下生长，冷冻并储存在液氮中。 我们希望从Semet取代的晶体（22 MET/103 kDa）收集一个完整的MAD数据集，以通过疯狂或悲伤的方法来解决相位问题。 此外，我们将收集高分辨率