Tomita T, Tokuhiro S, Hashimoto T, Aiba K, Saido T C, Maruyama K, Iwatsubo T
Department of Neuropathology and Neuroscience, Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo 113-0033, Japan.
J Biol Chem. 1998 Aug 14;273(33):21153-60. doi: 10.1074/jbc.273.33.21153.
Mutations in presenilin (PS) 1 or PS2 genes account for the majority of early-onset familial Alzheimer's disease, and these mutations have been shown to increase production of species of amyloid beta peptide (Abeta) ending at residue 42, i.e. the most amyloidogenic form of Abeta. To gain insight into the molecular mechanisms whereby mutant PS induces overproduction of Abeta42, we constructed cDNAs encoding mutant and/or truncated forms of PS2 and examined the secretion of Abeta42 from COS or neuro2a cells transfected with these genes. Cells expressing full-length PS2 harboring both N141I and M239V mutations in the same polypeptide induced overproduction of Abeta42, although the levels of Abeta42 were comparable with those in cells engineered to express PS2 with one or the other of these PS2 mutations. In contrast, cells engineered to express partially truncated PS2 (eliminating the COOH-terminal third of PS2 while retaining the endoproteolytic NH2-terminal fragment) and harboring a N141I mutation, as well as cells expressing COOH-terminal fragments of PS2, did not overproduce Abeta42, and the levels of Abeta42 were comparable with those in cells that expressed full-length, wild-type PS2 or fragments thereof. These data indicate that: (i) the Abeta42-promoting effects of mutant PS2 proteins reach the maximum level with a given single amino acid substitution (i.e. N141I or M239V); and (ii) the expression of full-length mutant PS2 is required for the overproduction of Abeta42. Hence, cooperative interactions of NH2- and COOH-terminal fragments generated from full-length mutant PS2 may be important for the overproduction of Abeta42 that may underlie familial Alzheimer's disease.
早老素(PS)1或PS2基因的突变是早发性家族性阿尔茨海默病的主要病因,这些突变已被证明会增加以42位残基结尾的淀粉样β肽(Aβ)的产生,即Aβ最具淀粉样变性的形式。为深入了解突变型PS诱导Aβ42过量产生的分子机制,我们构建了编码PS2突变体和/或截短形式的cDNA,并检测了转染这些基因的COS或neuro2a细胞中Aβ42的分泌情况。在同一多肽中同时携带N141I和M239V突变的全长PS2表达细胞会诱导Aβ42过量产生,尽管Aβ42的水平与经基因工程改造以表达其中一种PS2突变的细胞中的水平相当。相比之下,经基因工程改造以表达部分截短的PS2(去除PS2的COOH末端三分之一,同时保留蛋白水解后的NH2末端片段)并携带N141I突变的细胞,以及表达PS2的COOH末端片段的细胞,均未过量产生Aβ42,且Aβ42的水平与表达全长野生型PS2或其片段的细胞中的水平相当。这些数据表明:(i)突变型PS2蛋白促进Aβ42产生的作用在给定的单个氨基酸取代(即N141I或M239V)时达到最大水平;(ii)全长突变型PS2的表达是Aβ42过量产生所必需的。因此,全长突变型PS2产生的NH2末端和COOH末端片段之间的协同相互作用可能对Aβ42的过量产生很重要,而这可能是家族性阿尔茨海默病的基础。