Isolated nucleic acid molecule which codes for a 32 kDa protein having 11-CIS retinol dehydrogenase activity, and which associates with p63, a portion of a retinol binding protein receptor

Abstract

In accordance with this invention, an RPE cell membrane associated protein which has a molecular weight of about 32 kd, as determined by SDS-PAGE, has been discovered. This protein, referred to an “p32,” forms an oligomeric protein complex with the previously characterized p63 protein, a component of the membrane receptor for RBP. A nucleic acid molecule which codes for the p32 protein has also been isolated and sequence analysis shows that the p32 protein belongs to the family of short chain alcohol dehydrogenases, and exhibits 11-cis retinol dehydrogenase activity, the enzyme which catalyzes conversion of 11-cis-retinol into 11-cis retinaldehyde.

1 . Isolated nucleic acid molecule which codes for a mammalian protein having a molecular weight of about 32 kilodaltons as determined by SDS-PAGE, wherein said protein complexes with retinol binding protein receptor having a molecular weight of about 63 kilodaltons as determined by SDS-PAGE, wherein the complement of said isolated nucleic acid molecule hybridizes to the nucleotide sequence of SEQ ID NO:10 under stringent conditions.  
     
     
         2 . The isolated nucleic acid molecule of  claim 1 , wherein said mammalian protein is human protein.  
     
     
         3 . The isolated nucleic acid molecule of  claim 1 , wherein said mammalian protein is murine protein.  
     
     
         4 . The isolated nucleic acid molecule of  claim 1 , comprising cDNA.  
     
     
         5 . The isolated nucleic acid molecule of  claim 1 , comprising genomic DNA.  
     
     
         6 . The isolated nucleic acid molecule of  claim 2 , consisting of SEQ ID NO:14.  
     
     
         7 . The isolated nucleic acid molecule of  claim 2 , which encodes the amino acid sequence of SEQ ID NO:15.  
     
     
         8 . The isolated nucleic acid molecule of  claim 2 , which encodes the amino acid sequence of SEQ ID NO:15.  
     
     
         9 . The isolated nucleic acid molecule of  claim 3 , which encodes the amino acid sequence of SEQ ID NO:29.  
     
     
         10 . Expression vector comprising the isolated nucleic acid molecule of  claim 1 , operably linked to a promoter.  
     
     
         11 . Expression vector comprising the isolated nucleic acid molecule of  claim 2 , operably linked to a promoter.  
     
     
         12 . Cell line or bacterial cell strain, transformed or transfected with the expression vector of  claim 10 .  
     
     
         13 . Cell line or bacterial cell strain, transformed or transfected with the expression vector of  claim 11 .  
     
     
         14 . Isolated nucleic acid molecule which encodes a protein having retinol dehydrogenase activity, the complementary sequence of which hybridizes, under stringent conditions, to at least one nucleic acid molecule, selected from the group consisting of SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22 and SEQ ID NO:23, and sequences complementary thereto.  
     
     
         15 . The isolated nucleic acid molecule of  claim 14 , selected from the group consisting SEQ ID NOS:20, 21, 22 and 23.