Role of small molecule interactions and multiprotein assemblies in CYP1B1 disease-associated function and dysfunction

摘要 该项目的目标是确定突变、蛋白质水平升高和环境因素是如何影响基因表达的。 条件影响细胞色素P450 1B1（CYP1B1）的功能和功能障碍。添加 提出了一种仪器，以便于表征CYP1B1的小分子调节剂， 这一过程的代谢产物。CYP的选择性抑制是药物治疗的重要目标。 化学，但这些酶可以化学修饰其抑制剂，导致失活。的 要求高压液相色谱仪（HPLC），沿着用三个正交 检测系统，将允许准确和快速确定化学结构和酶 修改.这将用少量样品完成，因为分析物将来自 在组织培养中生长的细胞。由于CYP通常增加疏水底物的极性， 仪器包括分析水性介质所需的特定修改 解决方案这种溶液与大多数LC质谱仪不兼容。该设备将 也用于评估突变、蛋白质：蛋白质关联和环境条件是否改变了蛋白质的表达。 代谢产物，与缺氧条件下的组织选择性代谢的影响或由于 信号分子的存在。

Abstract The goal of this project is to determine how mutations, elevated protein levels, and environmental conditions impact the function and dysfunction of Cytochrome P450 1B1 (CYP1B1). The addition of instrumentation is proposed to facilitate characterization of small molecule modulators of CYP1B1, and the metabolic products of this CYP. Selective inhibition of CYPs is an important goal in medicinal chemistry, but these enzymes can chemically modify their inhibitors, resulting in deactivation. The requested High Pressure Liquid Chromatography (HPLC) instrument, along with three orthogonal detection systems, will allow for accurate and rapid determination of chemical structures and enzymatic modifications. This will be accomplished with small amounts of sample, as the analytes will come from cells grown in tissue culture. As CYPs generally increase the polarity of hydrophobic substrates, the instrumentation includes specific modifications that are required for analysis of aqueous media solutions. Such solutions are incompatible with most LC mass spectrometers. This equipment will be also used to evaluate if mutations, protein:protein association, and environmental conditions alter CYP metabolic products, with implications for tissue-selective metabolism under hypoxia or due to the presence of signaling molecules.