Andersen Olav M, Schmidt Vanessa, Spoelgen Robert, Gliemann Jørgen, Behlke Joachim, Galatis Denise, McKinstry William J, Parker Michael W, Masters Colin L, Hyman Bradley T, Cappai Roberto, Willnow Thomas E
Max-Delbrueck-Center for Molecular Medicine, Berlin, 13125 Berlin, Germany.
Biochemistry. 2006 Feb 28;45(8):2618-28. doi: 10.1021/bi052120v.
SorLA/LR11 is a sorting receptor that regulates the intracellular transport and processing of the amyloid precursor protein (APP) in neurons. SorLA/LR11-mediated binding results in sequestration of APP in the Golgi and in protection from processing into the amyloid-beta peptide (Abeta), the principal component of senile plaques in Alzheimer's disease (AD). To gain insight into the molecular mechanisms governing sorLA and APP interaction, we have dissected the respective protein interacting domains. Using a fluorescence resonance energy transfer (FRET) based assay of protein proximity, we identified binding sites in the extracellular regions of both proteins. Fine mapping by surface plasmon resonance analysis and analytical ultracentrifugation of recombinant APP and sorLA fragments further narrowed down the binding domains to the cluster of complement-type repeats in sorLA that forms a 1:1 stoichiometric complex with the carbohydrate-linked domain of APP. These data shed new light on the molecular determinants of neuronal APP trafficking and processing and on possible targets for intervention with senile plaque formation in patients with AD.
SorLA/LR11是一种分选受体,可调节神经元中淀粉样前体蛋白(APP)的细胞内运输和加工。SorLA/LR11介导的结合导致APP在高尔基体中被隔离,并防止其加工成β淀粉样肽(Aβ),Aβ是阿尔茨海默病(AD)中老年斑的主要成分。为了深入了解控制SorLA与APP相互作用的分子机制,我们剖析了各自的蛋白质相互作用结构域。通过基于荧光共振能量转移(FRET)的蛋白质接近度检测,我们在两种蛋白质的细胞外区域鉴定出结合位点。通过表面等离子体共振分析和重组APP与SorLA片段的分析超速离心进行精细定位,进一步将结合结构域缩小到SorLA中补体类型重复序列簇,该簇与APP的碳水化合物连接结构域形成1:1化学计量复合物。这些数据为神经元APP运输和加工的分子决定因素以及AD患者老年斑形成的可能干预靶点提供了新的线索。