STRUCTURAL ANALYSIS OF HUMAN MAIM PROTEIN

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 我们的目标是确定一种含有肌球蛋白激活相互作用基序(MAIM)的人类肌肉特异蛋白的结构，该基序与已知结构的蛋白质没有可识别的序列相似性。103 kDa蛋白已在大肠杆菌中得到表达和纯化，为可溶性活性蛋白。我们已经产生了天然蛋白质和Semet取代蛋白质的晶体。我们在我们的原产地(Rigaku MicroMax007，VariMax HR共焦光学系统)上收集了从天然晶体到2.8[CAPITAL_A_RING]分辨率的完整数据集，并进行了良好的统计。六方空间群(a=b=109.2[Capital_A_ring]，c=153.7[Capital_A_ring]，[Small_Alpha=[Small_beta]=90&#730；，[Small_Gamma]=120&#730；)的数据指数与我们从相对较小的晶体获得的强衍射值一致，其中一个分子位于不对称单元，溶剂含量为26%。SEMe取代的晶体具有与天然晶体相似的衍射性质，但已经长大到更大的尺寸。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(22Met/103 kDa)中收集完整的MAD数据集，以便用MAD或SAD方法解决相问题。此外，我们将收集一个高分辨率的

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