Differential effects of proteases involved in intracellular degradation of amyloid beta-protein between detergent-soluble and -insoluble pools in CHO-695 cells.

The deposition of amyloid beta-protein (A beta or beta A4) is a key feature of Alzheimer's disease. Most studies have focused on the generation of A beta, but little is known about the degradation of A beta. Recent reports suggest that insulin-degrading enzyme (IDE) and neutral endopeptidase (NEP) are involved in the extracellular degradation of A beta. To date, however, far less is known about the degradation of intracellular A beta. To elucidate the protease(s) responsible for the degradation of intracellular A beta, we investigated the effect of various protease inhibitors on A beta in two distinct intracellular pools (i.e., nonionic detergent-soluble and detergent-insoluble pools) in Chinese hamster ovary cells. Treatment with thiol and metal inhibitors resulted in the accumulation of intracellular A beta and oligomers in detergent-soluble and -insoluble fractions. The overexpression of thiol-metalloprotease IDE resulted in a marked reduction in levels of detergent-soluble intracellular A beta as well as extracellular A beta 40 and A beta 42. Moreover, intracellular A beta in the detergent-insoluble fraction extracted with 70% formic acid or 6 M guanidine hydrochloride decreased markedly in the cells overexpressing IDE. In contrast, expression of NEP degraded the A beta in the detergent-insoluble fraction markedly and partially degraded extracellular A beta 40 and A beta 42, but not intracellular soluble A beta. Thiorphan, an inhibitor of NEP, accumulated, albeit to a lesser extent, in insoluble A beta but not in soluble A beta. Thus, IDE appears to degrade intracellular A beta more effectively than does NEP in both the detergent-soluble and -insoluble fractions.