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.

项目概要 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、遗传漂变和迁移。了解这些力量影响的一个重要方法是， 特别是单个氨基酸改变突变对蛋白质功能的影响，是比较 我们的基因和蛋白质与我们最亲近的亲戚的基因和蛋白质的关系。在此过程中，选择模式（正 选择、负选择或放松约束）也可以推断。在过去的十年里，超过 数十个高质量的灵长类基因组序列已经发表，可以进行详细的研究 进化的力量作用于人类基因组和其他原始人类的基因组，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， 我们将通过使用以下方法进行定量功能测定来超越选择的计算预测 重组蛋白和合成肽来测量催化效率和底物的差异 人类精液中发现的高丰度细胞外蛋白的特异性。此类蛋白质常常被 通过计算预测为正选择、纯化选择和假生成的目标 在原始灵长类动物（人类和类人猿）中。 实验设计将包括测试人类重组蛋白的功能， 黑猩猩、大猩猩和猕猴。此外，我们将创建与最后一个相对应的蛋白质 人类和黑猩猩的共同祖先；人类、黑猩猩和大猩猩；和猕猴的 和原始人类，使用祖先序列重建。对于每个物种，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KLK3，以及前列腺TGM4的转谷氨酰胺酶活性。此外，效率 每种酶的特异性和特异性都将使用其天然底物（精囊）进行测试 表达 SEMG1 和 SEMG2，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，我们将检验物种在小酶活性方面存在差异的假设 来自 ACPP、SEMG1 和 SEMG2 的肽可形成淀粉样原纤维并增强 HIV 感染。 最后，我们将使用生物信息学方法来识别灵长类动物的基因，其进化可能是 由性选择或对性传播病毒的抵抗力驱动。 这项研究将通过让学生接触到杜肯大学的本科教育来改善 杰出的研究，同时显着改善 PI 实验室的研究环境， 部门和大学。