Mori H, Ishii K, Tomiyama T, Furiya Y, Sahara N, Asano S, Endo N, Shirasawa T, Takio K
Department of Molecular Biology, Tokyo Institute of Psychiatry.
Tohoku J Exp Med. 1994 Nov;174(3):251-62. doi: 10.1620/tjem.174.251.
Amyloid beta protein (A beta) in neuritic plaques of Alzheimer's disease has been found to be racemized and/or isomerized at their Asp residues. To elucidate the effect of racemization on the aggregation properties of A beta, we synthesized three kinds of A beta peptides in which D-Asp was substituted for L-Asp residues, i.e, normal A beta 1-40, [D-Asp7]A beta 1-40 and [D-Asp23]A beta 1-40. The aggregation and fibril formation of each peptide was examined by means of spectrofluorometry and electron microscopy. Of the three peptides, normal A beta showed the gradual increase of aggregation while [D-Asp7]A beta 1-40 and [D-Asp23] A beta 1-40 showed more enhanced aggregation at the final stage when the fibril formations were detected in all peptides solutions by electron microscopy. A comparative immunohistochemical study by anti-racemized A beta antibody and anti-A beta 1-42/43 antibody further showed the in vivo incorporation of D-Asp in senile plaques of Alzheimer's disease brains, which may be involved in plaque formation at the later stage than the deposition of the longer form of A beta (A beta 1-42/43). Taken together with the recent accumulated evidence on the aggregation mechanisms of A beta, the data presented here suggest that racemization may occur after the amyloid fibril formation but enhance the aggregation process by shifting the equilibrium of A beta from the soluble form to the insoluble form in Alzheimer's disease.
在阿尔茨海默病的神经炎性斑块中,已发现β淀粉样蛋白(Aβ)在其天冬氨酸(Asp)残基处发生了消旋化和/或异构化。为了阐明消旋化对Aβ聚集特性的影响,我们合成了三种Aβ肽,其中D - Asp取代了L - Asp残基,即正常的Aβ1 - 40、[D - Asp7]Aβ1 - 40和[D - Asp23]Aβ1 - 40。通过荧光光谱法和电子显微镜检查了每种肽的聚集和纤维形成情况。在这三种肽中,正常的Aβ显示出聚集的逐渐增加,而[D - Asp7]Aβ1 - 40和[D - Asp23]Aβ1 - 40在最终阶段显示出更强的聚集,此时通过电子显微镜在所有肽溶液中均检测到纤维形成。用抗消旋化Aβ抗体和抗Aβ1 - 42/43抗体进行的比较免疫组织化学研究进一步表明,D - Asp在阿尔茨海默病脑的老年斑中有体内掺入,这可能在比更长形式的Aβ(Aβ1 - 42/43)沉积更晚的阶段参与斑块形成。结合最近关于Aβ聚集机制积累的证据,此处呈现的数据表明,消旋化可能在淀粉样纤维形成后发生,但通过在阿尔茨海默病中将Aβ的平衡从可溶形式转变为不溶形式来增强聚集过程。