Protein turnover in pulmonary macrophages. Utilization of amino acids derived from protein degradation.
Conditions were defined under which rates of protein synthesis and degradation could be estimated in alveolar macrophages isolated from rabbits by pulmonary lavage and incubated in the presence of plasma concentrations of amino acids and 5.6 mM-glucose. Phenylalanine was validated as suitable precursor for use in these studies: it was not metabolized appreciably, except in the pathways of protein synthesis and degradation; it entered the cells rapidly; it maintained a stable intracellular concentration; and it was incorporated into protein at measurable rates. When extracellular phenylalanine was raised to a concentration sufficient to minimize dilution of the specific radioactivity of the precursor for protein synthesis with amino acid derived from protein degradation, the specific radioactivity of phenylalanyl-tRNA was only 60% of that of the extracellular amino acid. This relationship was unchanged in cells where proteolysis increased 2.5-fold after uptake and degradation of exogenous bovine serum albumin. In contrast, albumin prevented the decrease in phenylalanine incorporation observed in macrophages deprived of an exogenous source of amino acids. These observations suggested that macrophages preferentially re-utilized amino acids derived from the degradation of endogenous, but not from exogenous (albumin), protein. However, when the extracellular supply of amino acids was restricted, substrates derived from albumin catabolism could support the protein-synthetic pathway.
D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D066298:In Vitro Techniques; D008168:Lung / Q000166:cytology / Q000378:metabolism*; D008264:Macrophages / Q000378:metabolism*; D008297:Male; D010649:Phenylalanine / Q000378:metabolism; D011506:Proteins / Q000378:metabolism*; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism; D011817:Rabbits; D012710:Serum Albumin, Bovine / Q000378:metabolism
D000596:Amino Acids; D011506:Proteins; D012346:RNA, Transfer, Amino Acyl; D012710:Serum Albumin, Bovine; D010649:Phenylalanine