Sperm-specific AKAP protein genes and uses

Abstract

This invention provides isolated nucleic acid encoding sperm-specific protein kinase A anchoring proteins from mammalian species, recombinant expression constructs encoding said mammalian sperm-specific protein kinase A anchoring proteins, cells transformed with said constructs, and homogenous preparations of these proteins prepared using recombinant genetic techniques. The invention also provides analytic tools such as polyclonal antisera and monoclonal antibodies specific for said sperm-specific protein kinase A anchoring proteins of the invention. The invention also provides methods for isolating nucleic acid encoding mammalian sperm-specific protein kinase A anchoring proteins and recombinant genetic methods for producing said mammalian sperm-specific protein kinase A anchoring proteins of the invention. The invention particularly provides high throughput screening methods for screening compounds that can disrupt binding between protein kinase A and the sperm-specific protein kinase A anchoring protein and methods for determining and producing compounds, particularly antisense oligonucleotides, for inhibiting or preventing expression of sperm-specific protein kinase A anchoring protein in sperm, spermatids, or progenitor cells thereof. These methods are useful for development of effective, male-specific contraceptives, which are also provided by the invention.

We claim:  
     
         1 . A homogeneous composition of a mammalian sperm-specific protein kinase A anchoring protein having an apparent molecular weight of about 110 kildaltons.  
     
     
         2 . A homogeneous composition of a human sperm-specific protein kinase A anchoring protein according to  claim 1 , having an amino acid sequence identified by Seq. ID No. 16.  
     
     
         3 . A homogeneous composition of a bovine sperm-specific protein kinase A anchoring protein according to  claim 1 , having an amino acid sequence identified by Seq. ID No. 14.  
     
     
         4 . A homogeneous composition of a murine sperm-specific protein kinase A anchoring protein according to  claim 1 , having an amino acid sequence identified by Seq. ID No. 12.  
     
     
         5 . A method of screening a compound as an inhibitor of binding between a mammalian sperm-specific protein kinase A anchoring protein having an apparent molecular weight of about 110 kilodaltons and a regulatory subunit of protein kinase A, the method comprising the following steps: 
 (a) providing a first preparation of an isolated mammalian sperm-specific protein kinase A anchoring protein having an apparent molecular weight of about 110 kilodaltons;    (b) providing a second preparation of an isolated regulatory subunit of protein kinase A;    (c) contacting the first and second preparations under conditions permissible for binding of the protein kinase A anchoring protein and the regulatory subunit of protein kinase A in the presence or absence of the compound; and    (d) detecting a reduction in binding between the protein kinase A anchoring protein and the regulatory subunit of protein kinase A in the presence of the compound compared with binding in the absence of the compound.    
     
     
         6 . The method of  claim 5 , wherein either the protein kinase A anchoring protein or the regulatory subunit of protein kinase A is detectably-labeled.  
     
     
         7 . The method of  claim 5 , wherein the protein kinase A anchoring protein has an amino acid sequence identified by Seq. ID No. 16.  
     
     
         8 . The method of  claim 5 , wherein the protein kinase A anchoring protein has an amino acid sequence identified by Seq. ID No. 14.  
     
     
         9 . The method of  claim 5 , wherein the protein kinase A anchoring protein has an amino acid sequence identified by Seq. ID No. 12.  
     
     
         10 . The method of  claim 5 , wherein either the protein kinase A anchoring protein or the regulatory subunit of protein kinase A is covalently attached to a solid substrate.  
     
     
         11 . An antibody or fragment thereof that is immunologically reactive to a mammalian sperm-specific protein kinase A anchoring protein having an apparent molecular weight of 110 kilodaltons.  
     
     
         12 . The antibody or fragment thereof according to  claim 11 , wherein the antibody is a monoclonal antibody.  
     
     
         13 . The antibody or fragment thereof according to  claim 11 , wherein the protein kinase A anchoring protein is human sperm-specific protein kinase A anchoring protein having amino acid sequence identified by Seq. ID No. 16.  
     
     
         14 . The antibody or fragment thereof according to  claim 11 , wherein the protein kinase A anchoring protein is bovine sperm-specific protein kinase A anchoring protein having amino acid sequence identified by Seq. ID No. 14.  
     
     
         15 . The antibody or fragment thereof according to  claim 11 , wherein the protein kinase A anchoring protein is murine sperm-specific protein kinase A anchoring protein having amino acid sequence identified by Seq. ID No. 12.  
     
     
         16 . A cell line which produces an antibody or fragment thereof that is immunologically reactive to a mammalian sperm-specific protein kinase A anchoring protein having an apparent molecular weight of 110 kilodaltons.  
     
     
         17 . The cell line according to  claim 17 , wherein the antibody is a monoclonal antibody.  
     
     
         18 . The cell line according to  claim 17 , wherein the mammalian protein kinase A anchoring protein is human sperm-specific protein kinase A anchoring protein having amino acid sequence identified by Seq. ID No. 16.  
     
     
         19 . The cell line according to  claim 17 , wherein the mammalian protein kinase A anchoring protein is bovine sperm-specific protein kinase A anchoring protein having amino acid sequence identified by Seq. ID No. 14.  
     
     
         20 . The cell line according to  claim 17 , wherein the mammalian protein kinase A anchoring protein is murine sperm-specific protein kinase A anchoring protein having amino acid sequence identified by Seq. ID No. 12.  
     
     
         21 . An epitope of a mammalian protein kinase A anchoring protein is murine sperm-specific protein kinase A wherein the epitope is immunologically reactive to the antibody or fragment thereof according to  claim 11 .  
     
     
         22 . The epitope according to  claim 21  wherein the mammalian protein kinase A anchoring protein is human sperm-specific protein kinase A.  
     
     
         23 . The epitope according to  claim 21  wherein the mammalian protein kinase A anchoring protein is murine sperm-specific protein kinase A.  
     
     
         24 . The epitope according to  claim 21  wherein the mammalian protein kinase A anchoring protein is bovine sperm-specific protein kinase A.  
     
     
         25 . The epitope according to  claim 24  wherein the epitope has an amino acid sequence identified selected from the group consisting of Seq. ID Nos.: 1, 2, 3, 4, 5, 6, 7, 16 and 17.