QUANTITATIVE ANALYSIS OF AGE SPECIFIC VARIATION IN THE ABUNDANCE OF HUMAN FEMALE

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 人类唾液是一种富含蛋白质、易于获得的潜在局部和全身生物标志物来源，可用于监测病理条件下发生的变化;然而，人们对与正常衰老相关的丰度变化知之甚少。在这项研究中，我们进行了一个全面的蛋白质组分析收集唾液从腮腺健康女性受试者，分为两个年龄组1和2（20-30和55-65岁，分别）。在使用多维蛋白质鉴定技术（MudPIT）表征腮腺唾液之前，使用疏水电荷相互作用色谱法分离高丰度蛋白质和低丰度蛋白质。在两个年龄组中共鉴定出532种蛋白质。在这些蛋白质中，266种仅在一个年龄组中被鉴定，而266种蛋白质在两个年龄组中都有。在两个年龄组中鉴定的大多数蛋白质属于防御和免疫反应类别。值得注意的是，几种防御相关蛋白（例如，溶菌酶、乳铁蛋白和组胺素-1）在组2中显著更丰富。通过蛋白质印迹分析验证了选定的代表性质谱结果。我们的研究报告了第一次定量分析差异调节蛋白质的导管唾液收集年轻和老年女性受试者。这项研究支持使用高通量蛋白质组学作为一个强大的发现工具。这些结果为未来的研究提供了基础，以确定特定的唾液蛋白可能与特定于女性的年龄相关疾病有关。

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. Human saliva is a protein-rich, easily accessible source of potential local and systemic biomarkers to monitor changes that occur under pathological conditions; however, little is known about the changes in abundance associated with normal aging. In this study, we performed a comprehensive proteomic profiling of pooled saliva collected from the parotid glands of healthy female subjects, divided into two age groups 1 and 2 (20-30 and 55-65 years old, respectively). Hydrophobic charge interaction chromatography was used to separate high- from low-abundance proteins prior to characterization of the parotid saliva using multidimensional protein identification technology (MudPIT). Collectively, 532 proteins were identified in the two age groups. Of these proteins, 266 were identified exclusively in one age group, while 266 proteins were common to both groups. The majority of the proteins identified in the two age groups belonged to the defense and immune response category. Of note, several defense related proteins (e.g., lysozyme, lactoferrin and histatin-1) were significantly more abundant in group 2 as determined by G-test. Selected representative mass spectrometric findings were validated by Western blot analysis. Our study reports the first quantitative analysis of differentially regulated proteins in ductal saliva collected from young and older female subjects. This study supports the use of high-throughput proteomics as a robust discovery tool. Such results provide a foundation for future studies to identify specific salivary proteins which may be linked to age-related diseases specific to women.