The pleiotropic role of autophagy: from protein metabolism to bactericide
- N Mizushima
- Cell Death and Differentiation
Measurement of protein synthesis in individual organs is important in understanding metabolic changes in injury, sepsis, or starvation. Methods, mostly isotopic, for measuring synthesis are plagued by problems of experimental design and interpretation. Thus it is desirable to use a variety of methods based on different assumptions. The present study is the first to isolate radioactive aminoacyl-tRNA in the study of protein synthesis in muscle and skin. Male rats, 200-300 g, trained to eat chow for 4 hr/day were studied at 2 hr (absorptive) or 16 hr (postabsorptive) after a meal. Under ether anesthesia, a tracer dose of L-[4-5-3H(N)]-lysine was infused at a constant rate. At 20, 30, or 40 min 1 ml of arterial blood was withdrawn and 2-g samples of skin and thigh muscle were quickly excised and frozen. Samples were pooled from 4 to 7 rats for each infusion period. Concentrations and specific activities were determined for plasma lysine, and for free, tRNA, and protein-bound lysine in muscle and skin. Protein renewal rates in absorptive and postabsorptive periods averaged 6 and 9% per day in muscle, and 20 and 35% in skin. The data for muscle confirms results of other methods and suggests little contribution of rapidly turning over protein. The contribution of skin to whole body protein synthesis, about 500 mg . 100 g-1 . day-1, is similar in magnitude to the contributions of muscle, liver, or intestine.