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