Cryoelectron microscopy structure of purified gamma-secretase at 12 A resolution.
Cryoelectron microscopy structure of purified gamma-secretase at 12 A resolution.
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DOI：
10.1016/j.jmb.2008.10.078
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发表时间：
2009-01-16
影响因子：
5.6
通讯作者：
Li, Huilin
中科院分区：
文献类型：
--
作者：
Osenkowski, Pamela;Li, Hua;Ye, Wenjuan;Li, Dongyang;Aeschbach, Lorene;Fraering, Patrick C.;Wolfe, Michael S.;Selkoe, Dennis J.;Li, Huilin
γ-分泌酶（γ-Secretase）是一种完整的膜蛋白复合物，在神经元产生淀粉样β肽（Aβ）的过程中催化β-淀粉样前体蛋白（APP）的膜内裂解。因此，蛋白酶已成为开发治疗和预防阿尔茨海默病的药物的关键靶标。现有的生物化学研究对蛋白酶复合物的低聚组装状态存在冲突，并且其详细结构尚不清楚。在这里，我们报告，纯化的活性毛地黄皂苷中的人γ-分泌酶具有约230 kDa的总分子量时，通过扫描透射电子显微镜测量。这一结果支持了四种组分蛋白质中每一种都是单体的复合物。我们进一步报告了γ-分泌酶复合物在12 μ m分辨率下的三维结构，通过冷冻电镜和单粒子图像重建获得。该结构揭示了几个结构域的细胞外侧，三个溶剂可访问的低密度腔和一个潜在的底物结合的表面沟在跨膜区的复杂。
γ-Secretase, an integral membrane protein complex, catalyzes the intramembrane cleavage of the β-amyloid precursor protein (APP) during the neuronal production of the amyloid β-peptide (Aβ). As such, the protease has emerged as a key target for developing agents to treat and prevent Alzheimer's disease. Existing biochemical studies conflict on the oligomeric assembly state of the protease complex, and its detailed structure is not known. Here, we report that purified active human γ-secretase in digitonin has a total molecular mass of ~230 kDa when measured by scanning transmission electron microscopy. This result supports a complex that is monomeric for each of the four component proteins. We further report the 3-dimensional structure of the γ-secretase complex at 12 Å resolution, as obtained by cryo-EM and single particle image reconstruction. The structure reveals several domains on the extracellular side, three solvent-accessible low-density cavities and a potential substrate-binding surface groove in the transmembrane region of the complex.
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