Kinetics of degradation of "short-" and "long-lived" proteins in cultured mammalian cells

Abstract

Two distinct classes of protein referred to as short- and long-lived (Poole and Wibo, 1973) were found in pulse-chased HeLa S-3 and BHK 21/C13 cells. From experiments with pulse times ranging from 1 min to 20 h, a clear inverse correlation emerged between the pulse length and the percentage of protein which was hydrolysed intracellularly in the first h of chase. Using a 5 min pulse labelling with 3H-leucine, cells were either harvested immediately or after a 2 h chase. Approximately 35% of the label incorporated by cells was lost in a 2h chase; however, electrophoretic separation of cytosol and residual cytoplasmic fractions revealed no significant alteration in their protein profiles. The technique of selectively labelling "short" and "long-lived" proteins, which implies qualitative differences between then, is more readily interpreted as an artificial polarisation of a declining statistical probability curve of proteolysis with time which is similar for all nascent proteins.