Molecular basis of adaptation of seminal proteins of humans and other primates

人类和其他灵长类动物精蛋白适应的分子基础

• 批准号: 9305458
• 负责人: Michael Ignatius Jensen-Seaman
• 金额: $ 42.83万
• 依托单位: DUQUESNE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9305458
• 关键词: Affect; Alleles; Amino Acid Substitution; Amino Acids; Amyloid Fibrils; Bioinformatics; Biological Assay; Brain; Code; Complex; Computational Technique; Conflict (Psychology); Data; Data Analyses; Disease; Doctor of Philosophy; Environment; Enzymes; Evolution; Experimental Designs; Extracellular Protein; Fertilization; Frequencies; Gene Proteins; Genes; Genetic Drift; Genome; Genomics; Genotype; Goals; Gorilla gorilla; HIV; HIV Infections; Health; Hominidae; Human; Human Biology; Human Genome; Immune system; In Vitro; Individual; Investigation; KLK3 gene; Knowledge; Laboratories; Macaca; Measures; Medicine; Mentors; Modeling; Molecular; Molecular Evolution; Mutation; Natural Selections; Orthologous Gene; Pan Genus; Partner in relationship; Pattern; Peptide Fragments; Peptide Hydrolases; Peptides; Phenotype; Phosphoric Monoester Hydrolases; Physiological; Pongidae; Population Genetics; Preparation; Primates; Process; Prostate; Prostatic; Proteins; Publications; Publishing; Recombinant Proteins; Recording of previous events; Relaxation; Reproduction; Research; Resistance; SIV; Science; Seminal; Seminal Vesicles; Seminal fluid; Specificity; Students; Substrate Specificity; System; Taxonomy; Testing; Transglutaminases; Universities; Viral; Virus; Virus Diseases; base; career preparation; driving force; enzyme activity; extracellular; genetic evolution; improved; male; migration; pathogen; pressure; prostatic fraction Acid phosphatase isoenzyme; protein function; reconstruction; reproductive; response; species difference; sperm cell; synthetic peptide; tool; undergraduate education; undergraduate student

Project Summary The >20,000 protein-coding genes in our body have been shaped by the combined forces of mutation, natural selection, genetic drift, and migration. An essential way to understand the effects of these forces, especially the consequences of individual amino acid-changing mutations on protein function, is to compare our genes and proteins to those of our closest relatives. In doing so, the mode of selection (positive selection, negative selection, or relaxation of constraint) can also be inferred. Over the last decade, over a dozen high-quality primate genome sequences have been published, allowing for detailed investigation of the forces of evolution acting on the human genome and the genomes of other hominids, using complex models based on the principles of population genetics and molecular evolution. In the proposed research, we will go beyond computational predictions of selection by performing quantitative functional assays using recombinant proteins and synthetic peptides to measure differences in catalytic efficiency and substrate specificity of high-abundant extracellular proteins found in human semen. Such proteins have often been predicted computationally to be the targets of positive selection, purifying selection, and pseudogenization among the hominid primates (humans and the great apes). The experimental design will include testing the function of recombinant proteins from humans, chimpanzees, gorillas, and macaques. Furthermore, we will create the proteins corresponding to the last common ancestors of humans and chimpanzees; of humans, chimpanzees, and gorillas; and of macaques and the hominids, using ancestral sequence reconstruction. For each of these species, we will measure the phosphatase and peptidase activity of the prostatic acid phosphatase ACPP, the protease activity of the prostate expressed KLK3, and the transglutaminase activity of prostatic TGM4. Furthermore, the efficiency and specificity each of these enzymes will be tested using their natural substrates, the seminal vesicle expressed SEMG1 and SEMG2, to understand their coevolution. In addition to examining differences among species in enzyme activity, we will test the hypothesis that species differ in the ability of small peptides derived from ACPP, SEMG1, and SEMG2 to form amyloid fibrils and enhance HIV infection. Finally, we will use bioinformatics approaches to identify primate genes whose evolution may have been driven by either sexual selection or resistance to sexually transmitted viruses. This research will improve undergraduate education at Duquesne University by exposing students to meritorious research while significantly enhancing the research environment of the PI's laboratory, department, and university.

项目摘要 我们体内超过20，000个蛋白质编码基因是由突变的综合力量塑造的， 自然选择、遗传漂变和迁徙。理解这些力的影响的一个基本方法， 特别是单个氨基酸改变突变对蛋白质功能的影响， 我们的基因和蛋白质与我们最亲近的亲戚。在这样做时，选择模式（积极的 选择、否定选择或约束的放松）也可以被推断。在过去的十年里， 十几个高质量的灵长类动物基因组序列已经公布，允许详细调查， 进化的力量作用于人类基因组和其他原始人的基因组，使用复杂的 基于群体遗传学和分子进化原理的模型。在拟议的研究中， 我们将超越选择的计算预测，通过使用 重组蛋白和合成肽，以测量催化效率和底物的差异 在人类精液中发现的高丰度细胞外蛋白的特异性。这些蛋白质通常 预测计算的目标，积极的选择，净化选择，和假基因化 在原始灵长类动物（人类和类人猿）中。 实验设计将包括测试来自人类的重组蛋白的功能， 黑猩猩大猩猩和猕猴此外，我们将创建对应于最后一种蛋白质的蛋白质。 人类和黑猩猩的共同祖先;人类、黑猩猩和大猩猩的共同祖先;猕猴的共同祖先 和原始人类，使用祖先序列重建。对于每一个物种，我们将测量 前列腺酸性磷酸酶ACPP的磷酸酶和肽酶活性， 前列腺表达KLK 3和前列腺TGM 4的转氨酶活性。此外，效率 和特异性，这些酶中的每一种都将使用它们的天然底物， 表达SEMG 1和SEMG 2，以了解它们的共同进化。除了检查差异之外， 在酶活性的物种之间，我们将测试物种在小的能力上不同的假设， 来源于ACPP、SEMG 1和SEMG 2的肽，以形成淀粉样纤维并增强HIV感染。 最后，我们将使用生物信息学的方法来确定灵长类动物的基因，其进化可能是 由性选择或对性传播病毒的抵抗力驱动。 这项研究将改善迪克讷大学的本科教育， 在显著改善PI实验室的研究环境的同时， 系，大学。