Molecular Interactions in B cell Development

B 细胞发育中的分子相互作用

• 批准号: 7229988
• 负责人: JAMES D BALEJA
• 金额: $ 11.91万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-01-15 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7229988
• 关键词: Affinity; Amino Acids; Antibodies; Antibody Diversity; Antigens; Autoantibodies; Autoimmune Diseases; B-Cell Development; B-Lymphocytes; Binding; Calorimetry; Cell surface; Complex; Coupling; DNA-Protein Interaction; Development; Ensure; Future; Gene Rearrangement; Immunoglobulin Domain; Immunoglobulins; Individual; Knowledge; Laboratories; Light; Lipids; Maintenance; Measures; Methodology; Methods; Modeling; Molecular; Molecular Conformation; Mutagenesis; Nucleic Acid Binding; Peptides; Personal Satisfaction; Proteins; Reagent; Research Personnel; Role; Series; Signal Transduction; Staging; Structure; Surface Plasmon Resonance; Systemic Lupus Erythematosus; Techniques; Testing; Titrations; Work; immunoglobulin structure; innovation; insight; member; polypeptide; pre-B cell receptor; programs; protein function; protein protein interaction; research study; surrogate light chain; three dimensional structure

DESCRIPTION (provided by applicant): Maintenance of circulating antibodies requires proper B cell development. Gene rearrangement and editing provide for antibody diversity, but can also result in improperly formed antibody chains. There is a proofing mechanism at the pre-B cell stage of development in which a light-chain like molecule called the surrogate light chain probes the nascent heavy chain for proper folding. Successful complex formation results in assembly of the pre-B cell receptor that ensures survival and proliferation of that particular B cell. The mechanism by which the one and only surrogate light chain assembles and is able to pair with heavy chains is not known. The surrogate light chain comprises two polypeptides, VpreB and 14.1. The hypothesis that the two polypeptides of the surrogate light chain assemble to form a structure that resembles, in part, a mature light chain. Specific aim 1 is three-dimensional structure determination of the variable domain of the surrogate light chain. Using NMR methods we will determine the structure and identify the residues of the variable domain that interact with heavy chain. Specific Aim 2 focuses on identification of the role of unique, non-immunoglobulin domains in surrogate light chain assembly. The extent to which they participate in surrogate light chain assembly will be tested by measuring affinities of a series of constructs with and without the presence of the unique regions using isothermal titration calorimetry and surface plasmon resonance. The extent to which the unique, non-immunoglobulin domains of VpreB and 14.1 proteins have three-dimensional structure will also be determined. This project represents the first structural information for any component of the surrogate light chain of the pre-B cell receptor. Coupling structural information with other biophysical techniques that measure interactions is expected to provide insight on the mechanism of assembly. This R21 application is exploratory because the question is asked whether or not the unique regions have defined structure, either alone, or in complex with each other. Determining the structures will provide the necessary background for understanding and refining models put forward for signal transduction from the pre-B cell receptor that allow proper B cell formation and immunological competency.

描述（由申请人提供）：维护循环抗体需要适当的B细胞开发。基因重排和编辑提供了抗体多样性，但也可能导致形成不当的抗体链。在开发的前B细胞阶段有一个校对机制，其中一个称为替代光链的光链像分子一样探测新生的重链，以适当折叠。成功的复合形成会导致BED-B细胞受体组装，以确保该特定B细胞的存活和增殖。尚不清楚一个且唯一的替代光链组装并能够与重链配对的机制。替代光链包括两个多肽，即VPREB和14.1。替代光链的两个多肽组装以形成一种类似于成熟的轻链的结构的假设。特定目标1是替代光链可变域的三维结构测定。使用NMR方法，我们将确定结构并确定与重链相互作用的可变域的残基。特定目标2的重点是识别替代光链组件中独特的非免疫球蛋白结构域的作用。他们参与替代轻链组件的程度将通过测量有或不存在使用等温滴定量热法和表面等离子体共振的一系列构建体的亲和力来测试。 VPREB和14.1蛋白的独特的非免疫球蛋白结构域具有三维结构的程度。该项目代表了前B细胞受体的替代光链的任何组成部分的第一个结构信息。将结构信息与其他生物物理技术耦合，以测量相互作用的相互作用可以洞悉组装机理。此R21应用程序是探索性的，因为询问该问题是独特的区域是否单独或彼此复杂的结构。确定结构将为理解和精炼模型提供必要的背景，从而从前B细胞受体转导信号转导，从而使B细胞形成和免疫学能力适当。