AXEL BRUNGER PRT TIME

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。拟议的实验涉及对小蛋白质晶体的表征。所讨论的蛋白质是由两种不同蛋白质组成的相当大(~500 kDa)的复合体。在结晶紫染料的存在下，晶体呈蓝色，表明它们是由蛋白质组成的。然而，晶体只能以小平板的形式生长，平板表面的尺寸约为100-200微米，与平板垂直测量的尺寸为50微米。对晶体进行研究，以确定衍射度、孪生和马赛克。还将研究低温条件的潜在改善。这些晶体已经在各种不同的基质中生长。还将研究这些衬底对衍射性质的影响。确定空间群和收集这些晶体的原生集合将是这种光束时间转移的最终目标。Tavares-Maguk蛋白是组织膜结构域和蛋白质信号复合体的重要支架。该家族成员共有一个由PDZ结构域、SH3结构域和鸟苷酸激酶同源结构域组成的共同核心。已经确定了每个结构域的许多结合伙伴。涉及每个结构域的蛋白质-蛋白质相互作用解释了蛋白质在特定位置的聚集，例如在突触、隔膜和紧密连接以及极化上皮细胞的顶面和基面，以及膜信号通路在胚胎发育过程中诱导事件的能力。有人认为，分子内和分子间的相互作用调节着Maguk结构域的构象和结合活性。因此，我们有兴趣在分子水平上研究Maguk家族的第二个成员，以了解这些相互作用的结构基础和对伙伴结合的影响。

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. The proposed experiments involve the characterization of small protein crystals. The protein in question is a fairly large (~500 kDa) complex of two different proteins. The crystals stain blue in the presence of crystal violet dye, indicating that they are composed of protein. However, the crystals grow only as small plates, with dimensions of approximately 100-200 microns on the plate face and 50 microns measured orthogonal to the face. The crystals are to be studied to determine extent of diffraction, twinning and mosaicity. Potential improvements in cryo-conditions will also be examined. The crystals have been grown in a variety of different substrates. The effect of these substrates upon the diffraction properties will be studied as well. Determination of space group and the collection of a native set for these crystals would be the ultimate goal within this shift of beamtime. Tavares - MAGUK proteins are important scaffolds that organize membrane domains and protein signaling complexes. Members of the family share a common core composed of a PDZ domain, a SH3 domain and a guanylate kinase homology domain. Many binding partners for each of these domains have been identified. The protein-protein interactions involving each domain explain the clustering of proteins at specific sites, for instance at synapses, septate and tight junctions, and apical and basal surface of polarized epithelial cells, as well as the ability of membrane signaling pathways to induce events during embryonic development. It has been suggested that intra- and intermolecular interactions regulate conformation and binding activity of MAGUK domains. Therefore we are interested to study at molecular level two members of the MAGUK family in order to understand the structural basis of these interactions and implications for binding of partners.