Effect of protease inhibitors and clenbuterol on the in vitro degradation of dystrophin by endogenous proteases in human skeletal muscle.
The in vitro degradation of dystrophin protein by endogenous proteases in human skeletal muscle has been investigated using a tissue homogenate assay system with subsequent protein analysis via SDS polyacrylamide electrophoresis and immunoblotting (using a monoclonal antibody to the central rod region of dystrophin). The rate of dystrophin degradation and nature of the proteolytic fragments formed at pH 5.5 and pH 7.5 (corresponding to the two major protease groups of relevance to intracellular protein catabolism) were broadly similar; incorporation of protease inhibitors in the above system suggested that Ca2+ activated proteinase and cathepsin D are principally responsible for the degradation of dystrophin at pH 7.5 and pH 5.5 respectively. The rate of dystrophin degradation at pH 7.5 was reduced by approximately 20% in the presence of 10(-5) M clenbuterol, a beta-adrenoceptor agonist with therapeutic potential in the treatment of human muscle wasting diseases.
Maltin
C A
CA
Rowett Research Institute, Aberdeen, UK.
Jones
P
P
Mantle
D
D
D002402:Cathepsin D / Q000494:pharmacology; D002976:Clenbuterol / Q000494:pharmacology*; D016189:Dystrophin / Q000032:analysis / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000494:pharmacology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D009132:Muscles / Q000737:chemistry / Q000201:enzymology*; D011480:Protease Inhibitors / Q000494:pharmacology*
D016189:Dystrophin; D011480:Protease Inhibitors; D010450:Endopeptidases; D002402:Cathepsin D; D002976:Clenbuterol