Structure and Interactions of BDNF

BDNF 的结构和相互作用

• 批准号: 9042702
• 负责人: WILLIAM C BRACKEN
• 金额: $ 25.43万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9042702
• 关键词: Affinity; Antidepressive Agents; Anxiety Disorders; Binding; Biological; Biological Assay; Brain; Brain-Derived Neurotrophic Factor; Calorimetry; Cell Death; Cells; Cleaved cell; Complex; Elements; Episodic memory; Family member; Genes; Goals; Growth; Growth Cones; Growth Factor; Half-Life; Hippocampus (Brain); Human; Hydrogen; Label; Length; Lifting; Maintenance; Maps; Measurement; Measures; Memory; Memory impairment; Mental Depression; Methionine; Molecular; Molecular Conformation; N-terminal; Nerve Growth Factor Receptors; Neurodegenerative Disorders; Neuronal Differentiation; Neurons; Patients; Phosphorylation; Play; Population; Process; Protein Isoforms; Regulation; Research; Research Personnel; Risk; Role; Single Nucleotide Polymorphism; Site; Solutions; Structure; Synaptic Cleft; Synaptic plasticity; Techniques; Testing; Therapeutic; Titrations; Translating; Valine; Variant; Work; base; biophysical techniques; cell motility; design; in vivo; long term memory; meetings; neuronal growth; neuronal survival; neuropsychiatry; neurotrophic factor; neurotrophin 4; normal aging; prevent; protein folding; public health relevance; receptor binding; trafficking

 DESCRIPTION (provided by applicant): Brain derived neurotrophic factor (BDNF) stimulates the growth and survival of neuronal cells. Mild reductions in BDNF are associated with normal aging, memory impairment, neuropsychiatric and neurodegenerative disorders. Increased BDNF levels have been demonstrated to lift depression, prevent cell death and stimulate neuronal growth. In humans, a single nucleotide polymorphism in BDNF generates a valine to methionine substitution at residue 66 (Val66Met), located in the BDNF prodomain. The Met66 prodomain isoform is associated with memory impairment, depression and elevated risk of anxiety disorders. This substitution is found in 20% of the population, with 4% being homozygous. Common antidepressant therapies, that generally elevate BDNF levels, lose efficacy in patients bearing this isoform. We have recently observed that mature BDNF binds to its prodomain through interactions that directly involve the critical Val66Met site. In general, prodomain regions play important roles in protein folding, trafficking, stabilization, storage and regulation of mature growth factors. Specific isoform related differences in prodomain-BDNF interactions could provide a structural mechanism for the Met66 prodomain isoforms degenerative activity. We find that the isolated prodomain binds to mature BDNF with an affinity that suggests the complex could remain intact following secretion, potentially influencing BDNF activity. The goal of the proposed research is to structurally characterize this complex and elucidate critical differences in the Val66 and Met66 prodomain interactions with BDNF. To accomplish this goal we will pursue the following specific aims. Specific Aim 1: Produce and structurally characterize prodomain isoforms and mature BDNF growth factor. Specific Aim 2: Map the BDNF-prodomain binding interface and characterize differences in stability and function. We will pursue these aims using a combination of NMR and biophysical techniques combined with functional assessment of bioactivity using cell based assays. Many of the techniques applied in this study have not been directed toward the study of BDNF or other neurotrophin growth factors. We anticipate that this project will identify new molecular details about prodomain isoform interactions responsible for Val66Met differences in BDNF activity and function.

 描述（由申请人提供）：脑源性神经营养因子（BDNF）刺激神经元细胞的生长和存活。BDNF的轻度减少与正常衰老、记忆障碍、神经精神和神经退行性疾病有关。BDNF水平的增加已被证明可以解除抑郁症，防止细胞死亡和刺激神经元生长。在人类中，BDNF的单核苷酸多态性在位于BDNF前结构域的残基66（Val66Met）处产生缬氨酸至甲硫氨酸的取代。Met66前结构域同种型与记忆障碍、抑郁症和焦虑症风险升高相关。这种替换在20%的人群中发现，其中4%是纯合子。通常提高BDNF水平的常见抗抑郁疗法在携带这种亚型的患者中失去功效。我们最近观察到，成熟的BDNF通过直接涉及关键Val66Met位点的相互作用与其前结构域结合。一般来说，前结构域区域在成熟生长因子的蛋白质折叠、运输、稳定、储存和调节中起重要作用。前结构域-BDNF相互作用中的特异性同种型相关差异可以为Met66前结构域同种型变性活性提供结构机制。我们发现，分离的前结构域结合成熟的BDNF的亲和力，这表明该复合物可以保持完整的分泌后，潜在地影响BDNF的活性。拟议的研究的目标是从结构上表征这种复合物，并阐明Val66和Met66前结构域与BDNF相互作用的关键差异。为了实现这一目标，我们将努力实现以下具体目标。具体目标1：生产和结构表征前结构域异构体和成熟的BDNF生长因子。具体目标2：绘制BDNF-前结构域结合界面，并表征稳定性和功能的差异。我们将使用NMR和生物物理技术的组合，结合使用基于细胞的测定的生物活性的功能评估来追求这些目标。本研究中应用的许多技术并没有针对BDNF或其他神经营养生长因子的研究。我们预计，该项目将确定新的分子细节的前结构域异构体相互作用负责Val66Met的BDNF活性和功能的差异。