Antibacterial and anti-inflammatory activities of Parotid Secretory Protein

描述（由申请人提供）：该项目的长期目标是确定腮腺分泌蛋白（PSP；SPLUNC2）是否是口腔中的双重功能宿主防御蛋白。 PSP在唾液腺、口腔上皮细胞和唾液中表达，但其功能尚不清楚。 PSP 的表达受到细菌和 TNF 的上调，PSP 的计算机结构分析表明，它可能与杀菌/通透性增加蛋白 (BPI) 和 LPS 结合蛋白 (LBP) 相似。在这两种蛋白质中活性结构域的位置的指导下，设计了几种 PSP 肽，并发现它们在体外和体内均表现出抗菌和抗内毒素活性。基于 PSP 肽的这些初步结果和完整 PSP 的额外初步数据，假设检测到的 PSP 肽的双功能抗菌和抗炎活性反映在完整 PSP 蛋白的生物活性中。两个具体目标将测试 PSP 的拟议功能。目标 1 将表征 PSP 的抗菌活性。 PSP肽和重组PSP将用于生化、细胞生物学和体内实验，以确定PSP是否表现出抑菌或杀菌活性、细菌凝集和抑制生物膜形成。目标 2 将表征 PSP 的抗炎活性。 PSP 肽和重组 PSP 将用于生化、细胞生物学和体内实验，以确定 PSP 是否与 LPS 结合、在体外阻断 LPS 的作用并在 LPS 诱导的败血症模型中保护小鼠。如果得到证实，PSP 的拟议功能将扩大在口腔中表达并参与口腔粘膜防御的抗菌宿主防御蛋白组。因此，这里提出的研究将为未来旨在对抗感染和控制炎症的新临床方法的转化研究奠定基础。 公共健康相关性：该提案的目标是确定唾液蛋白腮腺分泌蛋白 (PSP) 的生物学功能。初步数据表明，PSP 在宿主防御口腔和呼吸道细菌中发挥作用。基于 PSP 序列的肽可能具有作为抗菌或抗炎成分的临床用途。

DESCRIPTION (provided by applicant): The long-term goal of this project is to determine if Parotid Secretory Protein (PSP; SPLUNC2) is a dual function host-defense protein in the oral cavity. PSP is expressed in salivary glands, oral epithelial cells and saliva but its function is unknown. Expression of PSP is up-regulated by bacteria and TNF and an in silico structural analysis of PSP revealed that it may be similar to bactericidal/permeability increasing protein (BPI) and LPS-binding protein (LBP). Guided by the location of active domains in these two proteins, several PSP peptides were designed and found to exhibit both antibacterial and anti-endotoxin activities in vitro and in vivo. Based on these initial results with PSP peptides and additional preliminary data for intact PSP it is hypothesized that the dual- function antibacterial and anti-inflammatory activities detected for PSP peptides are reflected in the biological activity of the intact PSP protein. Two specific aims will test the proposed functions of PSP. Aim 1 will characterize the antibacterial activity of PSP. PSP peptides and recombinant PSP will be used in biochemical, cell biological and in vivo experiments to determine if PSP exhibits bacteriostatic or bactericidal activity, bacterial agglutination, and inhibition of biofilm formation. Aim 2 will characterize the anti- inflammatory activity of PSP. PSP peptides and recombinant PSP will be used in biochemical, cell biological and in vivo experiments to determine if PSP binds LPS, blocks the action of LPS in vitro and protects mice in an LPS-induced sepsis model. If confirmed, the proposed functions of PSP will expand the group of antimicrobial host- defense proteins that are expressed in the oral cavity and participate in the defense of the oral mucosa. Thus, the research proposed here will lay the foundation for future translational research aimed at novel clinical approaches to combat infection and control inflammation. PUBLIC HEALTH RELEVANCE: The goal of this proposal is to identify the biological functions of the salivary protein Parotid Secretory Protein (PSP). Preliminary data suggest that PSP plays a role in host-defense against oral and airway bacteria. Peptides based on the PSP sequence may have clinical utility as antibacterial or anti-inflammatory components.