Wing S S, Chiang H L, Goldberg A L, Dice J F
Department of Cellular and Molecular Physiology, Harvard Medical School, Boston, MA 02115.
Biochem J. 1991 Apr 1;275 ( Pt 1)(Pt 1):165-9. doi: 10.1042/bj2750165.
In response to serum withdrawal, when overall rates of proteolysis increase in cultured fibroblasts, proteins containing peptide regions similar to Lys-Phe-Gln-Arg-Gln (KFERQ) are targeted to lysosomes for degradation, and the intracellular concentrations of these proteins decline [Chiang & Dice (1988) J. Biol. Chem. 263, 6797-6805]. To test whether such proteins are also selectively depleted in mammalian tissues in vivo, we have used affinity-purified polyclonal antibodies to KFERQ to detect proteins containing such sequences in tissues of fed and fasted rats. Immunoreactive cytosolic proteins were partially depleted from liver and heart of fasted rats, but the time course differed for these two tissues. Immunoreactive proteins in liver were lost during days 2 and 3 of fasting, whereas such proteins in heart were depleted within day 1 of fasting. In the same fasted rats, levels of immunoreactive cytosolic proteins did not change in two skeletal muscles, the dark soleus and the pale extensor digitorum longus. Immunoreactive proteins in a myofibrillar fraction were also partially depleted in heart, but not in skeletal muscles, of fasted rats. The most likely explanation for these results is that the protein loss in different tissues upon fasting results from selective activation of different proteolytic pathways. The increased proteolysis in liver and heart of fasted animals includes activation of the KFERQ-selective lysosomal pathway, whereas increased proteolysis in skeletal muscle does not.
血清撤除后,培养的成纤维细胞中蛋白水解的总体速率增加,含有类似于Lys-Phe-Gln-Arg-Gln(KFERQ)肽区域的蛋白质被靶向运送到溶酶体进行降解,这些蛋白质的细胞内浓度下降[蒋和戴斯(1988年)《生物化学杂志》263卷,6797 - 6805页]。为了测试这类蛋白质在哺乳动物组织体内是否也会被选择性消耗,我们使用了针对KFERQ的亲和纯化多克隆抗体来检测喂食和禁食大鼠组织中含有此类序列的蛋白质。禁食大鼠肝脏和心脏中的免疫反应性胞质蛋白部分减少,但这两个组织的时间进程有所不同。禁食第2天和第3天肝脏中的免疫反应性蛋白减少,而心脏中的此类蛋白在禁食第1天内就被消耗殆尽。在相同的禁食大鼠中,两块骨骼肌,即深色的比目鱼肌和浅色的趾长伸肌中免疫反应性胞质蛋白的水平没有变化。禁食大鼠心脏肌原纤维部分中的免疫反应性蛋白也部分减少,但骨骼肌中没有。对这些结果最可能的解释是,禁食时不同组织中的蛋白质损失是由不同蛋白水解途径的选择性激活导致的。禁食动物肝脏和心脏中蛋白水解增加包括KFERQ选择性溶酶体途径的激活,而骨骼肌中蛋白水解增加则不涉及该途径。