EAGER: Paleontological Proteomics Initiative: Developing Theory and Applications in Molecular Paleontology

EAGER：古生物学蛋白质组学计划：发展分子古生物学的理论和应用

• 批准号: 1547414
• 负责人: Ross MacPhee
• 金额: $ 28.38万
• 依托单位: American Museum Natural History
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2022-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1547414&HistoricalAwards=false
• 关键词: EAGER; Paleontological; Proteomics; Initiative; Developing

Molecular paleontology is a new field that uses information derived from biological molecules (biomolecules) to make inferences about evolutionary relationships, in this case for extinct organisms. During life, living things produce many kinds of biomolecules encoded by their DNA, and these may be preserved for varying lengths of time after the organism's death. Proteins, for example, are composed of amino acids, and each amino acid is specified by a precise piece of genetic information that varies slightly from species to species within an evolutionary group. Thus, by working out a protein's amino acid composition, the genetic sequence that originally produced it can be worked out indirectly, even in the absence of the DNA itself. This is significant for paleontological research because structural proteins like collagen, which make up almost all of the organic fraction of a bone, are very hardy and can last a long time after an animal's death; at least 4 million years in favorable circumstances, and possibly much longer. DNA, by contrast, degrades over a few tens to a few hundreds of thousands of years, even in the best preservational contexts. In recent years, instrumentation and lab techniques for acquiring compositional information from ancient biomolecules have greatly improved, enabling the researchers and their interdisciplinary collaborators in geochemistry to undertake a focused range of experiments in molecular paleontology. This research will advance the field of paleo-proteomics by addressing two main goals. The first goal will be to explore the limits of the technique and identify what kinds of fossils and what types of fossil preservation conditions yield the best results for analyses of biomolecules of extinct taxa. The second goal will apply the newly developed technique to specific 'test cases' to highlight the feasibility of the methods and their generality for application to diverse questions in systematics. Molecular methods have already proven vitally important for improving knowledge of the history of life on Earth, and the researchers' work will lead to both theoretical and practical improvements in ancient proteomics. How far back in time can ancient collagen proteomics actually reach and yield high-quality sequence information useful for phylogenetic studies? What are the best targets for preservation and systematic interpretation? Proof-of-concept investigations designed to answer these questions will focus on two areas of interest. (1) To establish fundamental geochemical boundary conditions affecting collagen survival, experiments will be conducted with a physically stabilized protein (collagen in fossil bone) and a proteome with restricted reactants (eggshell proteome). Work will center on assessing thermal age and controlling for mineral diagenesis, with analysis conducted via state-of-the-art mass spectrometry and diagenetic modeling to estimate the kinetics of key decay parameters (racemization, hydrolysis, oxidation, deamidation). (2) To establish the practical value of collagen proteomics for solving systematic problems, taxa from various temporal intervals in North and South America, West Indies, Asia, and Antarctica will be sampled for phylogenetic studies using the same instrumental approach as in (1). In addition to providing phylogenetically useful information, these investigations will further extend assessment of taphonomic conditions that enhance fossil protein preservation. The overall aim is to create a network of collaborative systematic paleontologists and protein geochemists interested in joint research in areas of mutual interest.

分子古生物学是一个新的领域，它使用来自生物分子的信息来推断进化关系，在这种情况下是灭绝的生物。在生命过程中，生物产生许多种由其DNA编码的生物分子，这些生物分子可能在生物体死亡后保存不同的时间长度。例如，蛋白质是由氨基酸组成的，每一种氨基酸都由一段精确的遗传信息指定，而在一个进化群体中，这段遗传信息在物种之间略有不同。因此，通过确定蛋白质的氨基酸组成，即使在没有DNA本身的情况下，也可以间接地确定最初产生蛋白质的遗传序列。 这对古生物学研究意义重大，因为构成骨骼几乎所有有机部分的胶原蛋白等结构蛋白非常哈代，在动物死亡后可以持续很长时间;在有利的环境下至少400万年，甚至可能更长。相比之下，即使在最好的环境中，DNA也会在几十到几十万年的时间里降解。近年来，从古代生物分子中获取成分信息的仪器和实验室技术有了很大的改进，使研究人员及其在地球化学领域的跨学科合作者能够在分子古生物学方面进行一系列集中的实验。这项研究将通过解决两个主要目标推进古蛋白质组学领域。第一个目标将是探索该技术的局限性，并确定哪种化石和哪种化石保存条件对分析灭绝分类群的生物分子产生最佳结果。第二个目标是将新开发的技术应用于特定的“测试案例”，以突出方法的可行性及其在系统学中应用于不同问题的一般性。分子方法已经被证明对于改善地球上生命历史的知识至关重要，研究人员的工作将导致古代蛋白质组学的理论和实践改进。古代胶原蛋白质组学实际上可以追溯到多远，并产生对系统发育研究有用的高质量序列信息？什么是保存和系统解释的最佳目标？为回答这些问题而设计的概念验证调查将侧重于两个感兴趣的领域。(1)为了建立影响胶原蛋白存活的基本地球化学边界条件，将用物理稳定的蛋白质（化石骨中的胶原蛋白）和具有限制反应物的蛋白质组（蛋壳蛋白质组）进行实验。工作将集中在评估热年龄和控制矿物成岩作用，通过最先进的质谱和成岩建模进行分析，以估计关键衰变参数（外消旋化，水解，氧化，脱酰胺）的动力学。(2)为了确定胶原蛋白质组学在解决系统问题方面的实用价值，将使用与（1）相同的仪器方法对来自南北美洲、西印度群岛、亚洲和南极洲的不同时间间隔的分类群进行采样，用于系统发育研究。除了提供生物学上有用的信息，这些调查将进一步扩大埋藏条件，提高化石蛋白质保存的评估。总体目标是建立一个合作的系统古生物学家和蛋白质地球化学家的网络，在共同感兴趣的领域进行联合研究。