STRUCTURAL ANALYSIS OF HUMAN MAIM PROTEIN

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 我们的目的是确定一个人肌肉特异性蛋白质的结构，含有肌球蛋白激活相互作用基序（MAIM），一个结构域与已知结构的蛋白质没有可识别的序列相似性。 从大肠杆菌中表达并纯化了103 kDa的蛋白。大肠杆菌作为可溶性活性蛋白。 我们已经产生了天然和SeMet取代蛋白质的晶体。 我们已经收集了完整的数据集，从原生晶体到2.8 [CAPITAL_A_RING]分辨率，在我们的原始源（Rigaku MicroMax 007 with VariMax HR共聚焦光学器件）上具有良好的统计数据。 六方空间群（a = B = 109.2 [CAPITAL_A_RING]，c = 153.7 [CAPITAL_A_RING]，[SMALL_ALPHA = [SMALL_BETA] = 90，[SMALL_Gamma] = 120）中的数据指数，其中不对称单元中有一个分子，溶剂含量为26%，与我们从相对较小的晶体中获得的强衍射一致。 SeMet取代的晶体具有类似于天然晶体的衍射性质，但已经生长到更大的尺寸。 晶体已经在低温条件下生长，冷冻，并储存在液氮中。 我们希望从SeMet取代的晶体（22 Met/103 kDa）收集完整的MAD数据集，用于通过MAD或SAD方法解决相位问题。 此外，我们将收集高分辨率

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