M tuberculosis PknB: Targeting the Extracellular domain

描述(由申请人提供)：结核病(TB)仍然是世界范围内，特别是在许多发展中国家的发病率和死亡率的主要原因。在艾滋病毒和结核病共同流行的地区，结核病是艾滋病毒感染者死亡的主要原因。在出现耐多药结核病的地方，包括在美国，治疗极其困难，死亡率很高，艾滋病毒/艾滋病患者受到的影响不成比例。需要新的预防和治疗方法，包括通过新靶点发挥作用的新药，以更有效地治疗药物敏感和耐多药结核病。由类真核细胞跨膜丝氨酸/苏氨酸激酶PKNA和PKNB介导的信号转导通路对结核分枝杆菌的生存至关重要。来自Pl实验室和结核分枝杆菌基因组序列的数据表明，这一信号通路调节细胞壁的合成和细胞分裂。R21建议的基本假设是，PKnB的胞外区(ED-PKNB)在体内通过与细胞外分子(S)的相互作用作为受体结构域或定位结构域发挥功能，这些相互作用对PKNB的功能是必不可少的。本研究的目的是通过鉴定和鉴定ED-PKNOB的合成配体来深入了解PKnA/B信号网络，并确定这些配体是否具有潜在的新型抗结核药物的先导作用。这些目标和方法将通过以下具体目标和方法来实现：1)使用新型玻片微阵列技术筛选与ED-PnuB结合的小分子化学文库，验证阳性结果并通过表面等离子体共振定量相互作用的强度，以及2)研究这些配体和活性衍生物的相互作用对分枝杆菌活力和细胞形态以及PnuB激酶活性的影响。这项研究的长期目标是深入了解PKnA/B信号通路，并确定ED-PKNOB的合成配体是否有潜力成为治疗或预防结核病的新方法的先导。

DESCRIPTION (provided by applicant): Tuberculosis (TB) remains a major cause of morbidity and mortality worldwide, particularly in much of the developing world. In regions where HIV and TB are co-prevalent, TB is a leading cause death in HIV-infected individuals. Where MDR-TB has emerged, including in the U.S., treatment is extremely difficult, mortality rates are high, and persons with HIV/AIDS have been disproportionately affected. New approaches to prevention and treatment, including new drugs that act through novel targets, are needed to treat drug- susceptible and MDR-TB more effectively. A signal transduction pathway mediated by the eukaryotic-like transmembrane serine/threonine kinases PknA and PknB is essential for the viability of M. tuberculosis. Data from the Pl's laboratory and the M. tuberculosis genome sequence indicate that this signaling pathway regulates cell wall synthesis and cell division. The underlying hypothesis of this R21 proposal is that the extracellular domain of PknB (Ed-PknB) functions in vivo as a receptor domain or as a localization domain, through interaction with extracellular molecule(s), and that these interactions are essential for the function of PknB. The goals of this research are to gain insight into the PknA/B signaling network through identification and characterization of synthetic ligands of ED-PknB, and to determine whether such ligands have potential as leads for novel anti-tuberculars. These goals will be addressed by the following specific aims and methods: 1) screening small molecule chemical libraries for compounds that bind to ED-PknB using a novel glass slide microarray technology, validating positives and quantifying the strength of interactions by surface plasmon resonance, and 2) investigating the effects of the interaction of these ligands, and active derivates, on mycobacterial viability and cell morphology, and on PknB kinase activity. The long-term goals of this research are to gain insight into the PknA/B signaling pathways, and to determine whether synthetic ligands of ED-PknB have potential as leads for new approaches to the treatment or prevention of TB.