The in Vivo brain interactome of the amyloid precursor protein
The in Vivo brain interactome of the amyloid precursor protein
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DOI：
10.1074/mcp.m700077-mcp200
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发表时间：
2008-01-01
影响因子：
7
通讯作者：
Schmitt-Ulms, Gerold
中科院分区：
文献类型：
--
作者：
Bai, Yu;Markham, Kelly;Schmitt-Ulms, Gerold
尽管进行了大量的研究，但淀粉样蛋白前体蛋白的生理功能和分子环境仍然是一个谜。在这里，我们描述了时间控制经心脏灌注交联的应用，这是一种在完整组织中蛋白质相互作用的体内作图方法，用于研究淀粉样前体蛋白（APP）的相互作用。为了深入了解所报道的蛋白质相互作用的特异性，该研究扩展到哺乳动物淀粉样蛋白前体样蛋白（APLP1和APLP2）。为了排除抽样偏差作为单个数据集差异的解释，我们对APP、APLP1和APLP2相互作用组进行了小规模的定量iTRAQ（相对和绝对定量等压标签）比较。相互作用组图证实了之前报道的8种APP相互作用，并揭示了在大脑中与APP空间接近的30多种其他蛋白质的身份。随后的验证研究证实了APP与富含亮氨酸的重复序列和含Ig结构域蛋白1之间的生理相互作用，证实了含Ig结构域蛋白1对APP蛋白水解加工的强烈影响，巩固了APP与p75在生物学上的相似性。
Despite intense research efforts, the physiological function and molecular environment of the amyloid precursor protein has remained enigmatic. Here we describe the application of time-controlled transcardiac perfusion cross-linking, a method for the in vivo mapping of protein interactions in intact tissue, to study the interactome of the amyloid precursor protein (APP). To gain insights into the specificity of reported protein interactions the study was extended to the mammalian amyloid precursor-like proteins (APLP1 and APLP2). To rule out sampling bias as an explanation for differences in the individual datasets, a small scale quantitative iTRAQ (isobaric tags for relative and absolute quantitation)-based comparison of APP, APLP1, and APLP2 interactomes was carried out. An interactome map was derived that confirmed eight previously reported interactions of APP and revealed the identity of more than 30 additional proteins that reside in spatial proximity to APP in the brain. Subsequent validation studies confirmed a physiological interaction between APP and leucine-rich repeat and Ig domain-containing protein 1, demonstrated a strong influence of Ig domain-containing protein 1 on the proteolytic processing of APP, and consolidated similarities in the biology of APP and p75.