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RUI: Aging & Biochemistry of a New Receptor Protein in Brain

RUI: Aging & Biochemistry of a New Receptor Protein in Brain
芮:老龄化
批准号:
8719741
负责人:
Lawrence Duffy
金额:
$10.73万
依托单位国家:
美国
项目类别:
Standard Grant
财政年份:
1988
资助国家:
美国
项目状态:
已结题
起止时间:
1988-03-15 至 1991-02-28

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中文摘要
翻译
这项研究是一个旨在了解脑受体蛋白分子老化的项目的初始阶段。达菲博士选择了一种受体蛋白,最近有报道称这种受体蛋白是在哺乳动物衰老的大脑中发现的淀粉样蛋白沉积的前体。其他人的初步研究表明,该基因在物种中高度保守,似乎在许多不同的细胞类型中表达,包括神经元。在这个初始阶段,达菲博士将调查各种细胞类型,包括内皮细胞、神经胶质细胞、神经元和成纤维细胞,以确定这种蛋白质的表达量是否足以进行生化研究。定量将通过使用为膜蛋白设计的现代FPLC程序分离蛋白质进行。该受体蛋白的纯化将随后使用抗体提高到人类蛋白质的各个区域,基于dna衍生的序列。达菲医生将通过生产针对纯化受体的单克隆抗体来补充这些抗体。当这些寻找合适的模型系统的研究正在进行时,Duffy博士将使用一系列肽类似物来研究淀粉样蛋白形成的必要元素,这些肽类似物可用于淀粉样蛋白片段的40个氨基酸。不同氨基酸对淀粉样蛋白结构的影响将使用电子显微镜、x射线和核磁共振方法进行监测。
英文摘要
This study is the initial phase in a project which seeks to understand the molecular aging of brian receptor proteins. Dr. Duffy has chosen a receptor protein recently reported to be the precursor to the amyloid deposits found in aging mammalian brains. Preliminary studies by others indicate that the gene is highly conserved across species and appears to be expressed in many different cell types, including neurons. In this initial phase, Dr. Duffy will survey a variety of cell types, including endothelial cells, glial, neurons and fibroblasts to see if the protein is expressed in sufficient quantities for biochemical studies. Quantitation will be performed by isolating the protein using modern FPLC procedures devised for membrane proteins. Purification of this receptor protein will be followed using antibodies raised to various regions of the human protein, based on the DNA-derived sequence. Dr. Duffy will supplement these antibodies by the production of monoclonal antibodies to the purified receptor. While these studies to find a suitable model system are in progress, Dr. Duffy will study the necessary elements of amyloid formation using a series of peptide analogues to the 40 amino acid amyloid fragment. The effect of different amino acids on the structure of amyloid will be monitored using EM, x-ray and NMR methodologies.
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