Cyclic GMP-binding, cyclic GMP-specific phosphodiesterase materials and methods

Abstract

The present invention provides novel purified and isolated nucleotide sequences encoding the cGMP-binding. cGMP-specific phosphodiesterase designated cGB-PDE. Also provided by the invention are methods and materials for the recombinant production of cGB-PDE polypeptide products and methods for identifying compounds which modulate the enzymatic activity of cGB-PDE polypeptides.

1 . A purified and isolated polynucleotide encoding cGB-PDE.  
     
     
         2 . The polynucleotide of  claim 1  which is a DNA sequence.  
     
     
         3 . The DNA sequence of  claim 2  which is a cDNA sequence or a biological replica thereof.  
     
     
         4 . The DNA sequence of  claim 2  which is a genomic DNA sequence or a biological replica thereof.  
     
     
         5 . An RNA transcript of the genomic DNA sequence of  claim 4 .  
     
     
         6 . The DNA sequence of  claim 2  which is a wholly or partially chemically synthesized DNA sequence or a biological replica thereof.  
     
     
         7 . The DNA sequence of  claim 4  further comprising an endogenous expression control DNA sequence.  
     
     
         8 . A DNA vector comprising a DNA sequence according to  claim 2 .  
     
     
         9 . The vector of  claim 8  wherein said DNA sequence is operatively linked to an expression control DNA sequence.  
     
     
         10 . A host cell stably transformed or transfected with a DNA sequence according to  claim 7  in a manner allowing the expression in said host cell of cGB-PDE polypeptide possessing a ligand/receptor binding biological activity or immunological property specific to cGB-PDE.  
     
     
         11 . A method for producing cGB-PDE polypeptide, said method comprising growing a host cell according to  claim 10  in a suitable nutrient medium and isolating cGB-PDE polypeptide from said cell or the medium of its growth.  
     
     
         12 . A polypeptide or peptide capable of specifically binding to cGB-PDE.  
     
     
         13 . An antibody substance according to  claim 12 .  
     
     
         14 . A monoclonal antibody according to  claim 13 .  
     
     
         15 . A hybridoma cell line producing a monoclonal antibody according to  claim 14 .  
     
     
         16 . A humanized antibody substance according to  claim 13 .  
     
     
         17 . An antisense polynucleotide specific for a polynucleotide encoding cGB-PDE.  
     
     
         18 . A DNA sequence encoding cGB-PDE and selected from the group consisting of: 
 (a) the DNA sequence set out in SEQ ID NO: 9 or 22;    (b) a DNA which hybridizes under stringent conditions to the DNA of (a); and    (c) a DNA sequence which, but for the redundancy of the generic code, would hybridize under stringent conditions to a DNA sequence of (a) or (b).    
     
     
         19 . A method for modulating the enzymatic activity of cGB-PDE, comprising contacting cGB-PDE with an effective amount of an agent that binds CGB-PDE and activates or inhibits cGB-PDE.  
     
     
         20 . The method of  claim 19  wherein the agent that binds cGB-PDE is selected from the group consisting of antibodies, peptides, proteins, oligonucleotides, antisense molecules, non-peptide compounds, and small molecules.  
     
     
         21 . The method of  claim 20 , wherein the agent that binds cGB-PDE is an anti-cGB-PDE antibody.  
     
     
         22 . A method for identifying an agent that specifically binds to cGB-PDE comprising: 
 (a) contacting cGB-PDE with an effective amount of a test agent; and    (b) determining if the test agent specifically binds cGB-PDE.    
     
     
         23 . A method for identifying an agent that specifically binds to cGB-PDE so as to modulate the enzymatic activity of cGB-PDE comprising: 
 (a) contacting cGB-PDE with an effective amount of a test agent;    (b) determining if the test agent specifically binds cGB-PDE; and    (c) testing for modulation of cGB-PDE activity.    
     
     
         24 . The method of  claim 23 , wherein the cGB-PDE is recombinant cGB-PDE.  
     
     
         25 . The method of  claim 23 , wherein the agent is selected from the group consisting of antibodies, peptides, proteins, oligonucleotides, antisense molecules, non-peptide compounds, and small molecules.  
     
     
         26 . A method of using an agent that modulates the enzymatic activity of cGB-PDE for treating a condition that involves signal transduction pathways utilizing cyclic nucleotides as second messengers, comprising administering to a subject an agent that modulates the activity of cGB-PDE.  
     
     
         27 . An agent identified by the method of  claim 24.