Sulfotyrosine, an essential determinant for diverse protein-protein interactions

磺基酪氨酸，多种蛋白质-蛋白质相互作用的重要决定因素

• 批准号: 10545692
• 负责人: PAMELA C RONALD
• 金额: $ 46.81万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-01 至 2028-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10545692
• 关键词: Address; Affinity; Agriculture; Animals; Antigen-Antibody Complex; Area; Awareness; Bacteria; Binding; Biological Assay; Biological Process; Cardiovascular Diseases; Cell surface; Cells; Coagulation Process; Complex; Development; Engineering; Foundations; Goals; HIV; Health; Human; Immune; Immune response; Knowledge; Ligand Binding; Medicine; Modification; Peptides; Phenotype; Plants; Play; Positioning Attribute; Process; Proteins; Reagent; Research; Role; Signal Transduction; Specificity; Spectrometry; Sulfate; Ticks; United States National Institutes of Health; Yeasts; blood treatment; gene discovery; insight; microorganism interaction; mutant; novel; protein protein interaction; receptor; response; therapeutic evaluation; tool; tyrosine O-sulfate; whole genome

1 This MIRA project will advance understanding of the function of tyrosine sulfation, a fundamental 2 modification that regulates diverse biological functions. For example, sulfation plays a critical role 3 in entry of HIV into human cells, in the interaction of microbial molecules with host immune 4 receptors and in the activity of tick-derived peptides, which hold potential for treatment of blood 5 clotting associated with cardiovascular diseases. Despite the increasing awareness of the 6 importance of tyrosine sulfation, most sulfated peptide complexes have not yet been fully 7 characterized. Further, we have only a partial view of the components essential for transducing 8 the response of such activated complexes. We also have limited strategies for efficiently 9 producing and testing the therapeutic potential of sulfated molecules. The PI is well positioned to 10 address these challenges. With support from the NIH, the PI discovered the host XA21 immune 11 receptor, a protein that is representative of a large class of plant and animal receptors. Further, 12 the PI's team isolated and characterized a sulfated peptide secreted by a Gram-negative 13 bacterium that binds to the XA21 receptor and activates the immune response. We propose 14 research in three related areas: We will isolate and characterize the diversity of receptors that 15 bind sTyr peptides, identify and analyze sTyr binding interfaces and pioneer a strategy to produce 16 novel sulfated molecules in an efficient manner. To accomplish our goals, we will employ new 17 experimental tools and approaches. These include use of established whole genome sequenced 18 mutant lines to rapidly assess phenotypes of newly discovered genes, sensitive assays to assess 19 ligand binding, state-of-the-art mass yeast cell surface display and spectrometry approaches to 20 engineer receptors and identify key components of immune complexes. The knowledge gained 21 from this research will provide new insights into an essential biological process, laying the 22 foundation for the development of reagents capable of inhibiting or activating cellular interactions 23 with a high degree of affinity and specificity with potential applications in research, agriculture and 24 medicine.

1这个MIRA项目将促进对酪氨酸硫酸化功能的理解，这是一个基本的 2修饰，调节不同的生物功能。例如，硫酸化作用起着关键作用， 3在艾滋病病毒进入人体细胞后，在微生物分子与宿主免疫相互作用中 4受体和蜱衍生肽的活性，其具有治疗血液 5与心血管疾病相关的凝血。尽管人们越来越意识到 6酪氨酸硫酸化的重要性，大多数硫酸化肽复合物尚未完全 7特征此外，我们对转导所必需的成分只有部分了解。 8、这种激活的复合物的反应。我们也有有限的策略， 9生产和测试硫酸化分子的治疗潜力。PI处于有利地位， 10应对这些挑战在NIH的支持下，PI发现宿主XA21免疫 11受体，一种代表一大类植物和动物受体的蛋白质。此外，本发明的目的是， PI的团队分离并表征了革兰氏阴性杆菌分泌的硫酸化肽， 13细菌结合到XA21受体并激活免疫反应。我们提出 14在三个相关领域的研究：我们将分离和表征受体的多样性， 15结合sTyr肽，鉴定和分析sTyr结合界面，并开创了一种产生 16种新型硫酸化分子。为了实现我们的目标，我们将采用新的 17个实验工具和方法。其中包括使用已建立的全基因组测序 18个突变株系，以快速评估新发现基因的表型，敏感的测定，以评估 19配体结合，最先进的大规模酵母细胞表面展示和光谱分析方法， 20工程受体和识别免疫复合物的关键成分。获得的知识 21这项研究将为一个重要的生物学过程提供新的见解， 22开发能够抑制或激活细胞相互作用的试剂的基础 23具有高度的亲和力和特异性，在研究、农业和 24药