Hamblet Natasha S, Ragland Brian, Ali Mervat, Conyers Barbara, Castora Frank J
Laboratory of Molecular Biochemistry, Department of Physiological Sciences, Eastern Virginia Medical School, Norfolk, VA 23507-1696, USA.
Electrophoresis. 2006 Feb;27(2):398-408. doi: 10.1002/elps.200500420.
A "mitochondrial hypothesis" of late onset Alzheimer's disease (AD) has been proposed. Biochemical studies indicate that there is a significant decrease in cytochrome oxidase (CO) activity as well as perturbed CO I and CO III mRNA levels in platelets and brain tissue from Alzheimer's patients. Using the electrophoretic mutation detection technique SSCP and DNA sequencing, we have identified 20 point mutations in the mitochondrial-encoded CO subunits (CO I, II, and III) in AD and age-matched control brain samples. Eight of the mutations are new variants of the mitochondrial genome. The efficiency of SSCP in detecting mutations in the CO subunits was estimated to be 80% when compared to dideoxy sequencing. One of the mutations (at position 9,861) results in a phenylalanine-->leucine substitution at a highly conserved residue in CO III. CO activity was reduced by an average of 35% in all AD brains compared to age-matched control samples, which agrees with previous reports. CO activity in one of the AD brain samples carrying the 9,861 mutation decreased by 80% relative to control brain samples, suggesting that the phenotypic expression of this mutation may result in reduced CO activity and compromised mitochondrial function.
一种关于晚发性阿尔茨海默病(AD)的“线粒体假说”已被提出。生化研究表明,阿尔茨海默病患者的血小板和脑组织中,细胞色素氧化酶(CO)活性显著降低,同时CO I和CO III mRNA水平也受到干扰。利用电泳突变检测技术单链构象多态性(SSCP)和DNA测序,我们在AD患者及年龄匹配的对照脑样本的线粒体编码CO亚基(CO I、II和III)中鉴定出20个点突变。其中8个突变是线粒体基因组的新变体。与双脱氧测序相比,SSCP检测CO亚基突变的效率估计为80%。其中一个突变(位于9861位)导致CO III中一个高度保守残基处的苯丙氨酸被亮氨酸取代。与年龄匹配的对照样本相比,所有AD脑样本中的CO活性平均降低了35%,这与之前的报道一致。携带9861突变的一个AD脑样本中的CO活性相对于对照脑样本降低了80%,表明该突变的表型表达可能导致CO活性降低和线粒体功能受损。