The proteasome inhibitors are used as research tools to study of the ATP-dependent ubiquitin-proteasome system. Some of them are at present undergoing clinical trials to be used as therapeutic agents for cancer or inflammation. These diseases are often accompanied by muscle wasting. We herein demonstrate findings about new proteasome inhibitors, belactosin A and C, and their direct effect on protein metabolism in rat skeletal muscle. M. soleus (SOL) and m. extensor digitorum longus (EDL) were dissected from both legs of male rats (40–60g) and incubated in a buffer containing belactosin A or C (30 μM) or no inhibitor. The release of amino acids into the medium was estimated using high performance liquid chromatography to calculate total and myofibrillar proteolysis. Chymotrypsin-like activity (CTLA) of proteasome and cathepsin B, L activity were determined by fluorometric assay. Protein synthesis and leucine oxidation were detected using specific activity of L-[1-14C] leucine added to medium. Inhibited and control muscles from the same rat were compared using paired t-test. The results indicate that after incubation with both belactosin A and C total proteolysis and CTLA of proteasome decreased while cathepsin B, L activity did not change in both SOL and EDL. Leucine oxidation was significantly enhanced in SOL, protein synthesis decreased in EDL. Myofibrillar proteolysis was reduced in both muscles in the presence of belactosin A only. In summary, belactosin A and C affected basic parameters of protein metabolism in rat skeletal muscle. The response was both muscle- and belactosin-type-dependent.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Amerik, A.Y. and Hochstrasser, M. (2004): Mechanism and function of deubiquitinating enzymes.Biochim Biophys Acta,1695, 189–207.
Anthony, J.C. Yoshizawa, F., Anthony, T.G., Vary, T.C., Jefferson, L.S. and Kimball, S.R. (2000): Leucine stimulates translation initiation in skeletal muscle of postabsorptive rats via a rapamycin-sensitive pathway.J Nutr,130, 2413–2419.
Asai, A., Tsujita, T., Sharma, S.V.et al. (2004): A new structural class of proteasome inhibitors identified by microbial screening using yeastbased assay.Biochem Pharmacol,67, 227–234.
Bailey, J.L., Wang, X., England, B.K., Price, S.R., Ding, X. and Mitch, W.E. (1996): The acidosis of chronic renal failure activates muscle proteolysis in rats by augmenting transcription of genes encoding proteins of the ATP — dependent ubiquitin — proteasome pathway.J Clin Invest,97, 1447–1453.
Cho, S.W., Romo, D. (2007): Total synthesis of (−)-belactosin C and derivatives via double diastereoselective tandem Mukaiyama aldol lactonizations.Org Lett,9, 1537–1540.
Cohen, S.A. and Michaud, D.P. (1993): Synthesis of a fluorescent derivatizing reagent, 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate, and its application for the analysis of hydrolysate amino acids via high-performance liquid chromatography.Anal Biochem,211, 279–287.
Du, J., Hu, Z. and Mitch, W.E. (2005): Molecular mechanisms activating muscle protein degradation in chronic kidney disease and other catabolic conditions.Eur J Clin Invest,35, 157–163.
Fang, C.H., Li, B.G., James, J.H.et al. (2005): Protein breakdown in muscle from burned rats is blocked by insulin-like growth factor and glycogen synthase kinase-3beta inhibitors.Endocrinology,146, 3141–3149.
Fang, C.H., Wang, J.J., Hobler, S., Li, B.G., Fischer, J.E. and Hasselgren, P.O. (1998): Proteasome blockers inhibit protein breakdown in skeletal muscle after burn injury in rats.Clin Sci,95, 225–233.
Fulks, R.M., Li, J.B. and Goldberg, A.L. (1975): Effects of insulin, glucose, and amino acids on protein turnover in rat diaphragm.J Biol Chem,250, 290–298.
Gaczynska, M., Rock, K.L. and Goldberg, A.L. (1993): Gamma-interferon and expression of MHC genes regulate peptide hydrolysis by proteasomes.Nature,365, 264–267.
Glickman MH, Ciechanover A. (2002): The ubiquitin-proteasome proteolytic pathway: destruction for the sake of construction.Physiol Rev,82, 373–428.
Glotzer, M., Murray, A.W. and Kirschner, M.W. (1991): Cyclin is degraded by the ubiquitin pathway.Nature,349, 132–138.
Gomes-Marcondes, M.C.C. and Tisdale, M.J. (2002): Induction of protein catabolism and the ubiquitin-proteasome pathway by mild oxidative stress.Cancer Lett,180, 69–74.
Groll, M., Bochtler, M., Brandstetter, H., Clausen, T. and Huber, R. (2005): Molecular machines for protein degradation.Chembiochem,6, 222–256.
Hobler, S.C., Tiao, G., Fischer, J.E., Monaco, J. and Hasselgren, P.O. (1998): Sepsis-induced increase in muscle proteolysis is blocked by specific proteasome inhibitors.Am J Physiol,274, R30–37.
Kadlcikova, J., Holecek, M., Safranek, R., Tilser, I. and Kessler, B.M. (2004): Effects of proteasome inhibitors MG132, ZL3VS and AdaAhx3 L3VS on protein metabolism in septic rats.Int J Exp Pathol,85(6): 365–371.
Kadlcikova, J., Holecek, M., Safranek, R. and Tilser I. (2005): Direct effects of proteasome inhibitor AdaAhx3L3VS on protein and amino acid metabolism in rat skeletal muscle.Physiol Res,54, 541–547.
Koohmaraie, M. and Kretchmar, D.H. (1990): Comparisons of four methods for quantification of lysosomal cysteine proteinase activities.J Anim Sci,68. 2362–2370.
Larionov, O.V. and de Meijere, A. (2004): Enantioselective total syntheses of belactosin A, belactosin C, and its homoanalogue.Org Lett,6(13), 2153–2166.
Lecker, S.H., Solomon, V., Mitch, W.E. and Goldberg A.L. (1999): Muscle protein breakdown and the critical role of the ubiquitin-proteasome pathway in normal and disease states.J Nutr,129, 227S-237S.
Lombardo, Y.B., Serdikoff, C., Thamotharan, M., Paul, H.S. and Adibi S.A. (1999): Inverse alterations of BCKA dehydrogenase activity in cardiac and skeletal muscles of diabetic rats.Am J Physiol,277, E685–692
Lowell, B.B., Ruderman, N.B. and Goodman, M.N. (1986): Regulation of myofibrillar protein degradation in rat skeletal muscle during brief and prolonged starvation.Metabolism,35, 1121–1127.
Lowry, O.H., Rosebrough, N.J., Farr, A.L. and Randall, R.J. (1951): Protein measurement with the Folin phenol reagent.J Biol Chem,193, 265–275.
Maizels, E.Z., Ruderman, N.B., Goodman, M.N. and Lau D. (1977): Effect of acetoacetate on glucose metabolism in the soleus and extensor digitorum longus muscles of the rat.Biochem J,162, 557–568.
Mayer, R.J. (2000): The meteoric rise of regulated intracellular proteolysis.Nat Rev Mol Cell Biol,1(2), 145–148.
Reverter, M., Lundh, T. and Lindberg, J.A. (1997): Determination of free amino acids in pig plasma by precolumn derivatization with 6-Nhydroxysuccinimidyl carbamate and high- performance liquid chromatography.J Chromatogr B Biomed Sci Appl,696(1): 1–8.
Rivett, A.J. (1993): Protesomes: multicatalytic proteinase complexes.Biochem J,291, 1–10.
Rock, K.L. and Goldberg A.L. (1999): Degradation of cell proteins and the generation of MHC class I-presented peptides.Annu Rev Immunol,17, 739–779.
Safranek, R., Holecek, M., Kadlcikova, J.et al. (2003): Effect of acute acidosis on protein and amino acid metabolism in rats.Clin Nutr,22(5), 437–443.
Salomon, V. and Goldberg, A.L. (1996): Importance of the ATP-ubiquitin-proteasome pathway in degradation of soluble and myofibrillar proteins in rabbit muscle extracts.J Biol Chem,271, 26690–26697.
Tardy, C., Autefage, H., García, V., Levade, T. and Andrieu-Abadie, N. (2004): Mannose 6-phosphorylated proteins are required for tumor necrosis factor-induced apoptosis: defective response in I-cell disease fibroblasts.J Biol Chem,279(51), 52914–52923.
Tawa, N.E. Jr., Odessey, R. and Goldberg, A.L. (1997): Inhibitors of the proteasome reduce the accelerated proteolysis in atrophying rat skeletal muscles.J Clin Invest,100(1), 197–203.
Thompson, M.G., Thom, A., Partridge, K.et al. (1999): Stimulation of myofibrillar protein degradation and expression of mRNA encoding the ubiquitin-proteasome system in C(2)C(12) myotubes by dexamethasone: effect of the proteasome inhibitor MG-132.J Cell Physiol,181(3), 455–461.
Tisdale, M.J. (2005): The ubiquitin-proteasome pathway as a therapeutic target for muscle wasting.J Support Oncol,3, 209–217.
Vary, T.C., Jefferson, L.S. and Kimball, S.R. (1999): Amino acid-induced stimulation of translation initiation in rat skeletal muscle.Am J Physiol,277, E1077–1086.
Wassner, S.J., Schlitzer, L. and Li, J.B. (1980): A rapid, sensitive method for the determination of 3-methylhistidine levels in urine and plasma using high-pressure liquid chromatography.Anal Biochem,104, 284–289.
About this article
Cite this article
Muthny, T., Kovarik, M., Sispera, L. et al. The effect of new proteasome inhibitors, belactosin A and C, on protein metabolism in isolated rat skeletal muscle. J Physiol Biochem 65, 137–146 (2009). https://doi.org/10.1007/BF03179064
- Skeletal muscle