ULTRA-HIGH RESOLUTION STUDIES OF CHOLESTEROL OXIDASE INCUBATED WITH XENON: HIV

与氙孵育的胆固醇氧化酶的超高分辨率研究：HIV

• 批准号: 7370450
• 负责人: S M SOLTIS
• 金额: $ 0.95万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-01 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7370450
• 关键词: ULTRA; RESOLUTION; STUDIES; CHOLESTEROL; OXIDASE

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. Ultra-high resolution structural studies of the flavoenzyme cholesterol oxidase (ChO) was carried out. ChO is a 55 kDa bifunctional flavoenzyme involved in the oxidation and isomerization of steroids that contain a 3-beta-hydroxyl group and a double bond at the C5 position of the steroid nucleus. The enzyme has been used clinically for the determination of serum cholesterol and most recently, due to the enzyme¿¿s ability to alter membrane structure, it has found applications as a potent larvicide. Moreover, the enzyme has also been proposed to be one of the virulence factors in Rhodococcus equi, an opportunistic pathogen in humans, associated with human immunodeficiency virus infection. In humans, Rhodococcus equi infection mainly affects the lungs with clinical pathological characteristics similar to pulmonary tuberculosis in immunocompromised patients. Cholesterol oxidase acts on eukaryotic membranes and its membrane damaging properties may be responsible for the observed cytotoxicity and macrophage destruction that accompanies Rhodococcal infection. Given the enzyme¿¿s prominent role in pathogenesis, we believe that a clear understanding of the substrate binding properties as well as the catalytic mechanism may aid in the development of inhibitors used as drugs to treat patients suffering from Rhodococcal infection. The purported channel for oxygen entry to the active site was studied using xenon incubation. Preliminary studies resulted in data sets extending to 0.85 ¿¿(cell dimensions: a=51.267, b=73.213 ¿¿ c=63.070 ¿¿ alpha=90.0 ¿¿ beta=105.132 ¿¿ gamma=90). Although xenon was found to bind, it was not in the purported channel.

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。对黄素酶胆固醇氧化酶（ChO）进行了超高分辨结构研究。ChO是一种55 kDa的双功能黄素酶，参与类固醇的氧化和异构化，类固醇在类固醇核的C5位置含有3-β-羟基和双键。该酶已在临床上用于测定血清胆固醇，最近，由于该酶的能力，改变膜结构，它已被发现作为一种有效的杀幼虫剂的应用。此外，该酶也被认为是马红球菌的毒力因子之一，马红球菌是一种人类机会致病菌，与人类免疫缺陷病毒感染有关。在人类中，马红球菌感染主要影响肺部，其临床病理特征类似于免疫功能低下患者的肺结核。胆固醇氧化酶作用于真核细胞膜，其膜损伤特性可能是红球菌感染时观察到的细胞毒性和巨噬细胞破坏的原因。鉴于酶？由于其在发病机制中的突出作用，我们相信，对底物结合特性以及催化机制的清楚理解可能有助于开发用作药物治疗红球菌感染患者的抑制剂。采用氙孵育法研究了氧进入活性部位的通道。初步研究得出的数据集扩展到0.85（细胞尺寸：a=51.267，B=73.213 c=63.070 α =90.0 β =105.132 γ =90）。虽然氙被发现绑定，它不是在所谓的通道。