Measurement of muscle protein degradation in live mice by accumulation of bestatin-induced peptides.
Bestatin, an aminopeptidase inhibitor, permits the degradation of cellular proteins to di- and tripeptides but interferes with the further breakdown of these peptides to amino acids. We propose to measure instant rates of protein degradation in skeletal muscles of intact mice by the accumulation of bestatin-induced intermediates. Muscle protein was labeled by injection of L-[guanidino-14C]arginine; 3 days later, maximum accumulation of intermediates was measured in abdominal wall muscles 10 min after the intravenous injection of 5 mg of bestatin. The peptides were partially purified and hydrolyzed in 6 N HCl, and the radioactivity in peptide-derived arginine was determined, after conversion to 14CO2 by treatment with arginase and urease. The measurement of bestatin-induced intermediates provides a unique tool for studying acute changes in muscle protein turnover in live mice. We observed a 62% increase in muscle protein breakdown after a 16-h fast, which was reversed by refeeding for 3.5 h, and a 38% increase after 3 days of protein depletion.
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Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire 03755-3844, USA.
Scornik
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D000009:Abdominal Muscles; D000818:Animals; D001120:Arginine / Q000378:metabolism; D001692:Biological Transport; D002250:Carbon Radioisotopes; D004435:Eating; D005215:Fasting; D007700:Kinetics; D007930:Leucine / Q000031:analogs & derivatives* / Q000493:pharmacokinetics / Q000494:pharmacology; D008297:Male; D051379:Mice; D009124:Muscle Proteins / Q000096:biosynthesis / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000187:drug effects / Q000378:metabolism*; D010446:Peptide Fragments / Q000032:analysis / Q000378:metabolism*; D011865:Radioisotope Dilution Technique; D014316:Tritium
D002250:Carbon Radioisotopes; D009124:Muscle Proteins; D010446:Peptide Fragments; D014316:Tritium; D001120:Arginine; D007930:Leucine; C012211:ubenimex