Analysis of cyclic nucleotide phosphodiesterase by isoelectric focusing coupled to a specific activity stain |
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Authors: | G Némoz A F Prigent H Pachéco |
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Institution: | Laboratoire de Chimie Biologique, INSERM U-205, Institut National des Sciences Appliquées de Lyon, Bâtiment 406, 20 avenue Albert Einstein, 69621 Villeurbanne Cédex, France |
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Abstract: | The procedure described allowed a convenient analysis of cyclic nucleotide phosphodiesterase. The different phosphodiesterase forms present in a crude cytosolic preparation from rat heart were separated by isoelectric focusing on a polyacrylamide gel plate. Phosphodiesterase activity bands were rendered evident by a specific staining method. They were then characterized by means of their substrate specificity and their sensitivity to selective phosphodiesterase inhibitors. The correspondence between the stain bands and the previously described activity peaks, obtained by a preparative technique and detected by radioisotopic enzyme assay, was also investigated. |
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Keywords: | cyclic nucleotide phosphodiesterase heart isoelectrofocusing histochemical stain isoenzyme analysis enzyme inhibitors |
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