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

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 金属和金属蛋白是许多细胞过程所必需的。我们实验室使用大分子X射线结晶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脱氧核糖核酸结合蛋白；自由基酶S-腺苷甲硫氨酸生物素合酶(BIOB)，参与生物素的生物合成；丙酮酸甲酸裂解酶(PflAE)，通过在PFL上形成一个蛋白质结合的甘氨酸基参与无氧代谢；以及羟丙基膦酸环氧酶(HppE)，它利用单核铁位置催化抗生素磷霉素的形成中独特的环氧化反应。我们已经解决并发表了所有这些蛋白质的初始晶体结构，其中一些工作是以前在SSRL进行的。我们现在正在寻求这些金属酶在多种状态下的进一步结晶学表征，以更全面地了解它们的功能性质。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. 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.