STRUCTURES OF NON-RIBOSOMAL PEPTIDE SYNTHETASES AND RELATED PROTEINS

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。 我们的实验室正在研究非核糖体多肽合成酶，这是一个合成抗生素和多肽铁载体的细菌蛋白质家族。这些蛋白质以模块化的方式工作，多个催化域连接到一个蛋白质中。这需要在蛋白质装配线上将共价结合的新生多肽从一个结构域传递到下一个结构域。 我们已经结晶了两个重要的复合体，其中一个是工程设计的两个结构域的嵌合结构，将允许表征结构域界面。我们还结晶了这两种蛋白质的复合体(不是经工程设计的嵌合蛋白质)，通过凝胶似乎在晶体中同时含有这两种蛋白质。这些晶体目前的衍射率只有5A，需要同步辐射才能找到最佳条件。(我们已经确定了其他晶体，包括PvdQ(来自假单胞菌的酰基酶)和琥珀酸半醛脱氢酶，以及嵌合复合体，因此我们不会花费全部时间进行筛选。)

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. Our lab is studying the non-ribosomal peptide synthetases, a family of bacterial proteins that synthesize antibiotics and peptide siderophores. These proteins operate in a modular fashion with multiple catalytic domains joined in a single protein. This requires a hand-off of the covalently bound nascent peptide from one domain to the next in the protein assembly line. We have crystallized two important complexes, one of an engineered two-domain, chimeric construct that will allow the characterization of the domain interface. We have also crystallized a complex of these two proteins (not engineered to be a chimeric protein) that appear by gels to have both proteins in the crystal. These crystals currently diffract to only 5A and require synchrotron radiation to find optimum conditions. (We have identified other crystals, including PvdQ (an acylase from Pseudomonas) and Succinate Semialdehyde Dehydrogenase, as well as the chimeric complex such that we will not spend the full time screening.)