Title: [62] Isolation and comparison of bovine heart cGMP-inhibited and cGMP-stimulated phosphodiesterases
Abstract: This chapter summarizes studies with two phosphodiesterase isozymes, the cGMP-stimulated and cGMP-inhibited phosphodiesterases that isolate from bovine cardiac muscle. Three different methods for producing stable, homogeneous preparations of these trace regulatory enzymes are described and a detailed comparison of the enzymes is presented. The chapter provides diagrammatic representations of the purification procedures. Procedure A enables one to purify both cGMP-stimulated and cGMP-inhibited phosphodiesterases from bovine cardiac muscle by taking advantage of the enzymes' differential binding to affinity resins. Procedure B utilizes isozyme-specific monoclonal antibodies to rapidly isolate cardiac muscle phosphodiesterase-antibody complexes for use in studies requiring preparations of catalytically active, nonproteolyzed enzymes. Procedure C represents a modification of the antibody isolation procedure B. This method results in homogeneous preparations of denatured but nonproteolyzed phosphodiesterases for use in a variety of protein structural studies. Although procedures B and C are described for purification of cGMP-inhibited phosphodiesterase, they can also be used to isolate cGMP-stimulated and Ca2+/calmodulin activated phosphodiesterases.
Publication Year: 1988
Publication Date: 1988-01-01
Language: en
Type: article
Indexed In: ['crossref', 'pubmed']
Access and Citation
Cited By Count: 11
AI Researcher Chatbot
Get quick answers to your questions about the article from our AI researcher chatbot