Biological Clocks: High Throughput Identification of Deterioration Markers and Dating of Museums' Proteinaceous Specimens

在化学部门和多学科活动办公室的化学测量和成像项目的支持下，莫伊尼博士和他在史密森学会的团队将解决追踪变质标记和测定文化遗产中蛋白质标本年代的需求。将开发具有高灵敏度、信息丰富的化学检测和低样品消耗的高通量分析技术，并对几类此类标记进行验证。为了证明这个概念，丝绸和羊毛纺织品，以及胶原蛋白和皮革标本将被用作博物馆蛋白质标本的例子。具体来说，本研究建议使用最先进的分离和质谱技术，研究在分子水平上跟踪蛋白质降解的三个因素——氨基酸外消旋化（AAR）、蛋白质脱酰胺化和蛋白质片段化（截断）。使用一系列年代准确的丝绸和羊毛纺织品，时间跨度大约从现在到2500年前，以及从现在到30000年前的胶原蛋白标本，这些退化的生物标记将被确定为一种可靠的年代测定方法。为了提高通量和减少样品消耗，将开发超快速毛细管电泳质谱法来取代传统的CE，将分析时间减少一个数量级至约一分钟。本项目开发的技术可能适用于其他博物馆的蛋白质类标本，包括油漆粘合剂、纸张、胶水、蛋白质类粘合剂等。开发可靠的生物钟具有广泛的实际应用，包括：食品工业中的食品安全和食品供应的生物测年；医院和医生办公室的疾病生物标志物检测；以及环境中生物污染物的检测。研究员将通过他的机构在当地的初中和高中开展推广项目和示范。这些活动的目标是让更广泛的社区参与到保护过程中来。研究生和研究生实习生，包括代表性不足的少数民族学生，将被鼓励在史密森学会开始他们的研究。

With support from the Chemical Measurement and Imaging Program in the Division of Chemistry and Office of Multidisciplinary Activities, Dr. Moini and his group at Smithsonian Institution will address the need for tracking deterioration markers and dating of proteinaceous specimens for cultural heritage purposes. High throughput analytical techniques with high sensitivity, information-rich chemical detection and low sample consumption will be developed, and several categories of such markers will be validated. For a proof of concept, silk and wool textiles, as well as collagen and leather specimens, will be used as examples of museums' proteinaceous specimens. Specifically, this proposal proposes to study the three factors that track protein degradation at a molecular level - amino acid racemization (AAR), protein deamidation, and protein fragmentation (truncation) - using state-of-the-art separation and mass spectrometry techniques. Using a series of well-dated silk and wool textiles, spanning in age from approximately present day to 2,500 years ago, and collagen specimens from present to ~30,000 BP, the biological markers of deterioration will be identified as a reliable means of dating. To increase throughput and reduce sample consumption, ultrafast capillary electrophoresis mass spectrometry will be developed to take the place of conventional CE, reducing analysis time by an order of magnitude to about one minute. The techniques developed in this project may be applicable to other museums' proteinaceous specimens, including paint binder, paper, glue, proteins-based adhesives, etc. The development of reliable biological clocks has a wide range of practical applications including: food safety and biological dating of food supplies in the food industry; disease biomarker detection in hospitals and doctors' offices; and detection of biological contaminants in the environment. The investigator will be engaged in outreach programs and demonstrations at local middle and high schools through his institution. The goal of these activities is to involve the broader community with the conservation process. Graduate and post graduate interns including underrepresented minority students will be encouraged to begin their research at the Smithsonian Institution.