CRYSTALLOGRAPHIC STUDIES OF METALLOPROTEINS (NIKR, BIOB, PFLAE, AND HPPE)

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 金属和金属蛋白对于许多细胞过程至关重要。 我们实验室利用高分子X射线晶体学来研究金属蛋白的结构和功能。 这些蛋白质包括镍调节转录因子 NikR，一种调节细胞镍摄取的 DNA 结合蛋白；自由基 S-腺苷甲硫氨酸 (SAM) 酶生物素合酶 (BioB) 参与生物素生物合成，丙酮酸甲酸裂解酶 (PFL) 激活酶 (PflAE) 通过在 PFL 上形成蛋白质结合的甘氨酰自由基参与无氧代谢；羟丙基膦酸环氧化酶（HppE），它利用单核铁位点催化独特的环氧化反应，形成抗生素磷霉素。我们已经解决并公布了所有这些蛋白质的初始晶体结构，其中一些工作之前是在 SSRL 进行的。我们现在正在寻求对这些金属酶在多种状态下的进一步晶体学表征，以更全面地了解它们的功能特性。

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. Metals and metalloproteins are essential for many cellular processes. Our laboratory uses macromolecular X-ray crystallography to study the structure and function of metalloproteins. These proteins include nickel regulatory transcription factor NikR, a DNA-binding protein that regulates cellular nickel uptake; the radical S-adenosylmethionine (SAM) enzymes biotin synthase (BioB), involved in biotin biosynthesis, and pyruvate formate-lyase (PFL) activating enzyme (PflAE), involved in anaerobic metabolism by formation of a protein bound glycyl radical on PFL; and hydroxypropylphosphonic acid epoxidase (HppE), which uses a mononuclear iron site to catalyze a unique epoxidation in the formation of the antibiotic fosfomycin. We have solved and published initial crystal structures of all of these proteins, and some of this work was carried out previously at SSRL. We are now in pursuit of further crystallographic characterization of these metalloenzymes in multiple states to more fully understand their functional properties.