Vargas J L, Knecht E, Grisolía S
Instituto de Investigaciones Citológicas de la Caja de Ahorros, Valencia, Spain.
Eur J Biochem. 1990 Feb 22;188(1):99-109. doi: 10.1111/j.1432-1033.1990.tb15376.x.
We have used a new approach to test the possible participation of lysosomes in the degradation of long-lived proteins. Rat liver lysosomal proteins were introduced, via multilamellar liposomes, into L-132 cells. Viability and protein synthesis were not impaired by this treatment. The liposomal content was released into the lysosomes of the cultured cells, as revealed by ferritin uptake and electron microscopy. Degradation rates of long-lived proteins increased with the uptake of lysosomal proteases. However, the increased protein degradation of chloroquine and leupeptin, in contrast to the inhibition by these reagents of the increased protein degradation of cells 'starved' of serum (step-down conditions). This approach opens a new way of investigating the degradation of intracellular proteins in cultured cells.
我们采用了一种新方法来测试溶酶体在长寿命蛋白质降解过程中可能的参与情况。通过多层脂质体将大鼠肝脏溶酶体蛋白导入L - 132细胞。这种处理并未损害细胞活力和蛋白质合成。如通过铁蛋白摄取和电子显微镜所显示的,脂质体内容物被释放到培养细胞的溶酶体中。长寿命蛋白质的降解速率随着溶酶体蛋白酶的摄取而增加。然而,氯喹和亮抑酶肽导致蛋白质降解增加,这与这些试剂对血清“饥饿”(逐步降低条件)细胞中蛋白质降解增加的抑制作用形成对比。这种方法为研究培养细胞中细胞内蛋白质的降解开辟了一条新途径。