The expression in porcine skeletal and cardiac muscle of calpastatin, the specific endogenous inhibitor of the calpain proteolytic system, was examined 16 h after a single dose of a specific beta(2)-agonist. Immunoblotting of extracts indicated that treatment increased skeletal calpastatin 135-kDa band intensity (P < 0.01), while in cardiac combined 145- and 135-kDa band intensity decreased (P < 0.05). Treatment increased skeletal (P < 0.01) but not cardiac calpastatin mRNA steady-state levels. Three types of cardiac calpastatin mRNA transcripts were identified by 5'-RACE. Types I and II encoded a putative XL region that originated either from exon 1x(A) or exon 1x(B), arranged in tandem. Type III predominated in skeletal muscle and originated from exon 1u, which was located 40-50 kb 3' to exons 1x(A) and 1x(B). The region 5' to exon 1u may act as an independent promoter regulated by a cAMP-dependent mechanisms, thereby explaining the differential response of calpastatin to adrenergic stimulation in cardiac and skeletal muscle.
Copyright 2001 Academic Press.