STRUCTURES OF NON-RIBOSOMAL PEPTIDE SYNTHETASES AND RELATED PROTEINS

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。子项目和 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 中心，不一定是研究者的机构。 我们实验室正在研究非核糖体肽合成酶，这是一个合成抗生素和肽铁载体的细菌蛋白质家族。这些蛋白质以模块化方式运作，多个催化结构域连接在单个蛋白质中。这需要在蛋白质装配线中将共价结合的新生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物，一个工程化的两个结构域，嵌合构建体，将允许域接口的表征。我们还结晶了这两种蛋白质的复合物（不是嵌合蛋白质），凝胶显示晶体中有两种蛋白质。这些晶体目前的折射率仅为5A，需要同步辐射来找到最佳条件。(We已经鉴定了其他晶体，包括PvdQ（来自假单胞菌的酰化酶）和琥珀酸半醛脱氢酶，以及嵌合复合物，使得我们将不花费全部时间筛选。

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.)