Pitfalls in invertebrate proteasome assays


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Reinhard.Saborowski [ at ] awi.de

Abstract

<jats:title>Summary</jats:title> <jats:p>The ubiquitin-proteasome system controls a variety of essential intracellular processes through directed protein turnover. The invertebrate proteasome has recently gained increasing interest with respect to central physiological processes and pathways in different taxa. A pitfall in proteasome-activity assays, represented by the trypsin-like, the chymotrypsin-like, or the caspase-like site, lies in the fact that most commonly-used experimental substrates are susceptible to degradation by non-proteasomal proteolytic enzymes, which can lead to erroneous interpretation of activity data obtained. Through the use of a proteasome-specific inhibitor, epoxomicin, we could show that the shares of proteasomal and non-proteasomal activities in the degradation of a model polypeptide substrate for the chymotrypsin-like activity vary considerably between invertebrate taxa. Crustacean muscle tissue and hemocytes showed almost exclusively proteasomal activity. In yeast, approximately 90% of total proteolytic activity can be attributed to the proteasome. In contrast, proteasomal activity comprises only 20-60% of the total proteolytic activity in bivalve tissues. These results reveal that, without verification of the shares of proteasomal and non-proteasomal activities in crude extracts through the use of highly specific inhibitors, common proteasomal enzyme assays should be used and interpreted with caution.</jats:p>



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Eprint ID
32733
DOI https://www.doi.org/10.1242/jeb.082792

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Götze, S. , Bose, A. , Abele, D. , Sokolova, I. and Saborowski, R. (2013): Pitfalls in invertebrate proteasome assays , Journal of Experimental Biology, 216 (8), pp. 1351-1354 . doi: https://www.doi.org/10.1242/jeb.082792


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