Differential effects of detergents, fatty acids, cations and heating on ostrich skeletal muscle 20S proteasome.
The 20S proteasome, the catalytic core of the 26S proteasome, has previously been isolated, purified and partially characterised from ostrich skeletal muscle (Thomas, A.R., Oosthuizen, V., Naude, R.J., Muramoto, K. 2002. Biol. Chem. 383, 1267-1270). Due to the apparent latency of the 20S proteasome purified from various sources, this study focuses on further characterising the ostrich enzyme in terms of the effects of selected detergents, fatty acids and cations, as well as heating at 60 degrees C, on four of its activities. Results showed that ostrich skeletal muscle 20S proteasome was affected in a non-concentration-dependent manner by the selected detergents and fatty acids. Monounsaturated fatty acids, unlike unsaturated fatty acids, showed no major effects on the activities of the ostrich enzyme. The enzyme did not show sensitivity towards monovalent cations and the only divalent cations that showed a relevant effect were Ca2+ and Mg2+. Heating at 60 degrees C for 1-2 min had a substantial activating effect only on the peptidylglutamylpeptide-hydrolase (PGPH) and caseinolytic activities. In conclusion, many of the effects by the abovementioned reagents and conditions were noticeably different to those shown on different sources of the enzyme, further demonstrating the unique kinetic characteristics of the ostrich skeletal muscle 20S proteasome.
Thomas
Adele R
AR
Department of Biochemistry and Microbiology, University of Port Elizabeth, PO Box 1600, Port Elizabeth 6000, South Africa.
Oosthuizen
Vaughan
V
Naudé
Ryno J
RJ
D000818:Animals; D002412:Cations / Q000494:pharmacology*; D003902:Detergents / Q000494:pharmacology*; D004795:Enzyme Stability; D005227:Fatty Acids / Q000494:pharmacology*; D006361:Heating; D018482:Muscle, Skeletal / Q000201:enzymology*; D046988:Proteasome Endopeptidase Complex / Q000737:chemistry / Q000187:drug effects*; D020310:Struthioniformes / Q000378:metabolism*; D013379:Substrate Specificity
D002412:Cations; D003902:Detergents; D005227:Fatty Acids; D046988:Proteasome Endopeptidase Complex