DNA molecule encoding a variant alpha-2B-adrenoaceptor protein and uses thereof

Abstract

This invention relates to a DNA sequence comprising a nucleotide sequence encoding a variant α 2B -adrenoceptor protein and to the variant α 2B -adrenoceptor protein as well as a method for screening a subject to determine if the subject is a carrier of a variant gene that encodes the variant α 2B -adrenoceptor protein. This invention also relates to a method for treating a mammal suffering from vascular contraction of coronary arteries, the method comprising the step of administering a selective α 2B -adrenoceptor antagonist to the mammal. This invention further relates to transgenic animals comprising a human DNA molecule encoding human α 2B -adrenoceptor protein or the variant α 2B -adrenoceptor protein.

1 . A nucleic acid encoding a variant C2B-adrenoceptor protein, the variant protein comprises a deletion of at least 1 glutamate from a glutamic acid repeat element of 12 glutamates, amino acids 298-309, in an acidic stretch of 18 amino acids 294-311, located in a 3 rd  intracellular loop of the receptor protein.  
     
     
         2 . The nucleic acid of  claim 1 , wherein the variant protein comprises a deletion of 3 glutamates, amino acids 307-309, from said glutamic acid repeat element of 12 glutamates, amino acids 298-309, in said acidic stretch of 18 amino acids 294-311, located in the 3 rd  intracellular loop of the receptor polypeptide.  
     
     
         3 . The nucleic acid of  claim 2  comprising a genomic nucleotide sequence as set forth in SEQ ID NO:1.  
     
     
         4 . The nucleic acid of  claim 1 , wherein said nucleic acid is cDNA.  
     
     
         5 . An RNA sequence fully complementary to the DNA sequence of  claim 1 .  
     
     
         6 . A variant α 2B -adrenoceptor protein comprising a deletion of at least 1 glutamate from said glutamic acid repeat element of 12 glutamates, amino acids 298-309, in said acidic stretch of 18 amino acids 294-311, located in the 3 rd  intracellular loop of the receptor polypeptide.  
     
     
         7 . The variant α 2B -adrenoceptor protein of  claim 6 , wherein the protine comprises a deletion of 3 glutamates, amino acids 307-309, from said glutamic acid repeat element of 12 glutamates, amino acids 298-309, in said acidic stretch of 18 amino acids 294-311, located in the 3 rd  intracellular loop of the receptor polypeptide.  
     
     
         8 . The variant α 2B -adrenoceptor protein of  claim 7  comprising an amino acid sequence set forth in SEQ ID NO: 2.  
     
     
         9 . An assay for determining the presence or absence of the nucleic acid of  claim 1 .  
     
     
         10 . The assay of  claim 9 , wherein the assay is a DNA-assay.  
     
     
         11 . A method for determining the presence or absence in a biological sample of the nucleic acid of  claim 1  comprising contacting a single-stranded form of said nucleic acid if present in the sample with a capturing nucleic acid probe and a detector nucleic acid probe to form a complex and detecting the presence or absence of the complex.  
     
     
         12 . The method of  claim 11 , wherein the capturing nucleic acid probe is attached or capable of attaching to a solid phase, and comprises a cDNA encoding the variant α 2B -adrenoceptor protein, wherein a detected signal from the solid phase is an indication of the presence in the sample of said nucleic acid.  
     
     
         13 . The method according to  claim 11 , wherein the capturing nucleic acid probe is attached or capable of attaching to a solid phase, and comprises a cDNA encoding a non-variant α 2B -adrenoceptor protein, wherein a detected signal from the solid phase is an indication of the absence in the sample of said nucleic acid.  
     
     
         14 . A method for screening a subject to determine if the subject is at risk for developing a disease involving vascular contraction of coronary arteries or is in need of α 2B -selective or α 2B -nonselective α 2 -adrenoceptor antagonist therapy, said method comprises obtaining a biological sample of the subject and determining whether the subject (i) has the insertion/insertion (I/I) or deletion/insertion (D/I) genotypes of the human α 2B -adrenoceptor protein or (ii) has the D/D genotype of the human α 2B -adrenoceptor protein, wherein if the subject has the D/D genotype, the subject is at risk for developing a disease involving vascular contraction of coronary arteries or is in need of α 2B -selective α 2 -adrenoceptor antagonist therapy.  
     
     
         15 . The method of  claim 14 , wherein the assay is a DNA-assay.  
     
     
         16 . A capturing probe which comprises a single strand of the cDNA of  claim 4 .  
     
     
         17 . A capturing probe which comprises a single strand of a cDNA encoding a non-variant α 2 B-adrenoceptor protein.