期刊
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS
卷 427, 期 1, 页码 8-15出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.abb.2004.04.001
关键词
calpain; calpastatin; promoter analysis; skeletal muscle; cardiac muscle
资金
- Biotechnology and Biological Sciences Research Council [D20453] Funding Source: Medline
- Biotechnology and Biological Sciences Research Council [D20453] Funding Source: researchfish
Calpastatin is the specific endogenous inhibitor of calpain proteinase that is encoded by a single gene. Transient transfection assays in both a non-fusing skeletal muscle and non-muscle cell-line demonstrated that the putative porcine calpastatin promoter regions 5' to exons 1xa, 1xb, and 1u were functional. Both real-time quantitative and semi-quantitative RT-PCR on porcine skeletal muscle total RNA indicated that steady-state expression of Type I and III mRNAs containing exons 1xa and 1u, respectively, was at equivalent levels whilst the expression of Type 11 mRNA containing exon 1xb was significantly less (p < 0.001). Immunoprobing of Western blotted muscle extracts with an antibody raised against a peptide sequence encoded by exon 1xa indicated that Type I protein was expressed and that there was significantly more Type I protein in cardiac than skeletal muscle (p < 0.001). The results suggest that the expression of the single calpastatin gene was differentially controlled at several levels. (C) 2004 Elsevier Inc. All rights reserved.
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