Alterations in components of the ubiquitin-protein ligase system following maturation of reticulocytes to erythrocytes.
Previous studies have shown that the activity of the ubiquitin-mediated proteolytic system declines markedly following reticulocyte maturation, but the specific alterations responsible for this phenomenon have not been defined. We find that the rate of ATP-dependent degradation of 125I-albumin is reduced 20-fold in lysates of rabbit erythrocytes, as compared to reticulocyte lysates. The activity of the proteolytic system in erythrocyte extracts can be restored by supplementation with components of the ubiquitin-protein ligase system purified from reticulocytes by affinity chromatography. These components are the ubiquitin-carrier protein E2, the activity of which is nearly completely absent, and the ligase E3, the activity of which is partially reduced in erythrocytes. Erythrocyte extracts contain other ligases which attach a single, or a few ubiquitin molecules to proteins; these products are different from the multi-ubiquitin derivatives which are formed by the ligase system of protein breakdown. Mature red cells may thus serve to distinguish between different ubiquitin-protein ligase systems with presumably different functions.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002846:Chromatography, Affinity; D004229:Dithiothreitol; D004912:Erythrocytes / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D008025:Ligases / Q000097:blood*; D011817:Rabbits; D012156:Reticulocytes / Q000201:enzymology*; D044767:Ubiquitin-Protein Ligases
D000255:Adenosine Triphosphate; D044767:Ubiquitin-Protein Ligases; D008025:Ligases; D004229:Dithiothreitol