SGER: Phylogenetic and Indigeneity Implications of MALDI-MS Determined Osteocalcin Protein Sequences

SGER：MALDI-MS 测定的骨钙素蛋白序列的系统发育和本土性影响

• 批准号: 9614235
• 负责人: Peggy Ostrom
• 金额: $ 1万
• 依托单位: Michigan State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-09-01 至 1997-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9614235&HistoricalAwards=false
• 关键词: SGER; Phylogenetic; Indigeneity; Implications; MALDI

9614235 Ostrom A new approach, matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS), to determine the amino acid sequence of the protein osteocalcin in bones of ancient organisms and their closest modern ancestors to establish preliminary evolutionary relationships and assess indigeneity. The use of osteocalcin, the particular samples available, and the application of MALDI-MS provide a unique position of strength for the proposed research. Osteocalcin has several extremely important attributes which include intimate association with the hydroxyapatite mineral phase of bone, preferential preservation over time, exclusive occurrence in vertebrate mineralized tissues, absence in common contaminators including microbes, invertebrates and plants, retention of immunological specificity via radioimmunoassay (RIA), and the existence of an extensive database of protein sequences for osteocalcin. In our particular case, prior work has confirmed the presence of osteocalcin via RIA for all ancient samples. The MALDI-MS technique is not subject to the myriad of problems that confront traditional methods for protein sequencing. Although this approach has never been attempted on ancient proteins, we intend to demonstrate its potential to delineate ancient protein sequences and, thus, identify indigenous organic matter. Given the exclusive yet ubiquitous occurrence of osteocalcin among vertebrates, the techniques developed in this project will be directly applicable for molecular studies on any number of other vertebrate lineages with obscured evolutionary relationships.

小行星9614235 一种新的方法，基质辅助激光解吸电离质谱法（MALDI-MS），以确定蛋白质骨钙素的氨基酸序列在古代生物及其最近的现代祖先的骨骼建立初步的进化关系和评估individuality。 骨钙素的使用、可获得的特定样品以及MALDI-MS的应用为所提出的研究提供了独特的优势。 骨钙素具有几个极其重要的属性，包括与骨的羟基磷灰石矿物相的紧密结合，随时间优先保存，仅存在于脊椎动物矿化组织中，不存在包括微生物、无脊椎动物和植物在内的常见污染物，通过放射免疫测定（RIA）保留免疫特异性，以及存在广泛的骨钙素蛋白质序列数据库。 在我们的特殊情况下，先前的工作已经证实了骨钙素的存在，通过RIA的所有古代样本。 MALDI-MS技术不受传统蛋白质测序方法所面临的无数问题的影响。 虽然这种方法从来没有尝试过古老的蛋白质，我们打算证明其潜力描绘古老的蛋白质序列，从而确定土著有机物质。 鉴于骨钙素在脊椎动物中的独特而普遍的发生，本项目中开发的技术将直接适用于任何数量的其他脊椎动物谱系的分子研究与模糊的进化关系。