Investigating Molecular Assembly and Dynamics of Tick Cement proteins

总结 蜱虫必须用它们带倒钩的口器（hypostome）深深刺入宿主的真皮层 （皮肤），然后包住hypostome在水泥锥，以保持坚定，疼痛和瘙痒- 自由附着，随后将传染病病原体传播给受害者。两种类型 在蜱虫中发现了大量的水泥;早期的水泥在几分钟后分泌出来， 附着并迅速硬化，附着后24小时分泌的晚期骨水泥 并逐渐变硬。这些分泌的水泥锥是由几个 富含甘氨酸的蛋白质（GRP）是一个大家族的不同蛋白质，已知其 粘合和拉伸特性。这些特性可能在宿主皮肤中起重要作用 附着以在长时间内获得不间断的血液供应。我们长久以来- 在区域拨款申请的长期计划是揭示的功能和生化特性， 骨水泥锥组件、隐形附件、噬血（供血）和 气候适应我们将测试我们的中心假设，即生物化学和生物力学 GRP的属性有助于未检测到的对主机的连接，提供对 通过用作水泥锥装置的支架来不间断地供应血液。我们 多学科团队将使用人工膜喂养， 结构、蛋白质组学和反向遗传学方法。具体目标是：1）确定 独特的唾液GRP在骨水泥锥组装和使用骨水泥锥固定中的作用 人工膜进料系统，和2）定义未表征的生化特性 重组表达的水泥锥蛋白。靶向蜱粘固蛋白可以提供 创造一种不利于蜱虫附着的环境，并最终中断 将蜱传病原体接种到宿主体内。了解水泥的作用机制 组装，让蜱安全稳定，隐形附件将提供洞察 开发新的控制和预防方法。

Summary Ticks must use their barbed mouthparts (hypostome) to pierce deeply into the host’s dermis (skin), then encase the hypostome in a cement cone in order to maintain a firm, pain and itch- free attachment, subsequently transmitting infectious disease agents into the victim. Two types of cement have been found in ticks; early cement which is secreted only minutes after attachment and hardens rapidly, and late cement which is secreted 24 hours after attachment to the host and hardens gradually. These secreted cement cones are composed of several glycine-rich proteins (GRPs) which are a large family of diverse proteins that are known for their adhesive and tensile characteristics. Such properties may play an important role in host skin attachment to gain an uninterrupted supply of blood over a prolonged period of time. Our long- term plan in the AREA grant application is to reveal the functions and biochemical properties of novel GRPs in cement cone assembly, stealth attachment, hematophagy (blood feeding) and climatic adaptation. We will test our central hypothesis that biochemical and biomechanical properties of GRPs assist in the undetected attachment to the host, providing access to an uninterrupted supply of blood by serving as scaffolding for the cement cone apparatus. Our multidisciplinary team will test the hypothesis using a combination of artificial membrane feeding, structural, proteomics, and reverse genetics approaches. Specific aims are to: 1) determine the role of unique salivary GRPs in cement cone assembly and secure attachment by using an artificial membrane feeding system, and 2) define the biochemical properties of uncharacterized recombinantly expressed cement cone proteins. Targeting tick cement proteins may provide an innovative way to create a hostile situation for tick attachment, and ultimately interrupt the inoculation of tick-borne pathogens into the host. Understanding the mechanisms of cement assembly that allow the tick to secure a stable, stealthy attachment will provide insight into developing new control and prevention methods.