Pacemake ion channel proteins and uses thereof

Abstract

The present invention provides an isolated nucleic acid encoding a BCNG protein or a portion thereof or BCNG-related protein or a portion thereof. The present invention further provides a method for identifying a nucleic acid in a sample which encodes a BCNG protein or a BCNG-related protein. The present invention also provides a method for testing whether a compound affects the expression of a BCNG protein or a BCNG-related protein. In addition, the present invention further provides a method for identifying a compound capable of interacting with a BCNG protein or a BCNG-related protein. Also, the present invention provides a method for identifing a compound capable of modulating BCNG protein or BCNG-related protein activity. Further, the present invention also provides a method of treating a condition in a subject which comprises administering to the subject an amount of the provided compound, effective to treat the condition. Finally, the present invention provides a pharmaceutical composition which comprises the provided compound and a pharmaceutically acceptable carrier.

What is claimed is:  
     
         1 . An isolated nucleic acid encoding a BCNG protein or a portion thereof.  
     
     
         2 . An isolated nucleic acid encoding a BCNG-related protein or a portion thereof.  
     
     
         3 . The nucleic acid of  claim 1 , wherein the BCNG protein is encoded by the sequence shown in mBCNG-1 (ATCC Accession No. ______) (Seq.ID.No.:1), mBCNG-2 (Seq.ID.No.:5), mBCNG-3 (Seq.ID.No.:9), mBCNG-4 (Seq.ID.No.:11), hBCNG-1 (ATCC Accession No. ______) (Seq.ID.No.:3) or hBCNG-2 (Seq.ID.No.:7).  
     
     
         4 . The nucleic acid of  claim 1  or  2 , wherein the nucleic acid is DNA or RNA.  
     
     
         5 . The nucleic acid of  claim 1  or  2 , wherein the nucleic acid is cDNA.  
     
     
         6 . The nucleic acid of  claim 5 , wherein the cDNA has the nucleotide sequence shown in SEQ. ID. No.: 1 for mBCNG-1 (ATCC Accession No. ______), SEQ. ID. No.: 3 for hBCNG-1, SEQ. ID. No.: 5 for mBCNG-2, SEQ. ID. No.: 7 for hBCNG-2, SEQ. ID. No.: 9 for mBCNG-3, or SEQ. ID. No.:11 for mBCNG-4.  
     
     
         7 . A vector comprising the nucleic acid of  claim 1  or  2 .  
     
     
         8 . The vector of  claim 7 , wherein the vector comprises viral or plasmid DNA.  
     
     
         9 . An expression vector comprising the nucleic acid of  claim 1  or  2  and regulatory elements.  
     
     
         10 . A host vector system which comprises the expression vector of  claim 9  in a suitable host.  
     
     
         11 . A vector, which comprises cDNA encoding mBCNG-1 (ATCC Accession No. ______).  
     
     
         12 . A vector, which comprises cDNA encoding hBCNG-1 (ATCC Accession No. ______).  
     
     
         13 . The host vector system of  claim 10 , wherein the suitable host is a bacterial cell, a eukaryotic cell, a mammalian cell or an insect cell.  
     
     
         14 . An isolated BCNG protein.  
     
     
         15 . An isolated BCNG-related protein.  
     
     
         16 . The protein of  claim 14 , wherein the BCNG protein has substantially the same amino acid sequence shown in Seq.ID.No.:2 for mBCNG-1 (FIG. 8A), Seq.ID.No.:6 for mBCNG-2. (FIG. 8A), Seq.ID.No.:10 for mBCNG-3 (FIG. 8A), Seq.ID.No.12 for mBCNG-4 (FIG. 8A), Seq.ID.No.:4 for hBCNG-1 (FIG. 8A), Seq.ID.No.:8 for hBCNG-2 (FIG. 8A) or a portion thereof.  
     
     
         17 . The protein of  claim 15 , wherein the BCNG-related protein has substantial homology to the amino acid sequence shown in Seq.ID.No.:2 mBCNG-1 (FIG. 8A), Seq.ID.No.:6 for mBCNG-2 (FIG. 8A), Seq.ID.No.:10 for mBCNG-3 (FIG. 8A), Seq.ID.No.12 for mBCNG-4 (FIG. 8A), Seq.ID.No.:4 for hBCNG-1 (FIG. 8A), Seq.ID.No.:8 for hBCNG-2 (FIG. 8A) or a portion thereof.  
     
     
         18 . A composition comprising a nucleic acid, encoding a BCNG protein or a BCNG-related protein or a portion thereof and a carrier.  
     
     
         19 . The composition of  claim 18 , wherein the nucleic acid comprises substantially the same coding sequence as the coding sequence shown in SEQ. ID. No.: 1 for mBCNG-1, SEQ. ID. No.: 0.3 for hBCNG-1, SEQ. ID. No.: 5 for mBCNG-2, SEQ. ID. No.: 7 for hBCNG-2, SEQ. ID. No.: 9 for mBCNG-3, SEQ. ID. No.:11 for mBCNG-4 or portion thereof.  
     
     
         20 . A composition comprising a BCNG protein or a BCNG-related protein or portion thereof and a carrier.  
     
     
         21 . The composition of  claim 20 , wherein the BCNG protein or BCNG-related protein comprises substantially the same amino acid sequence as the amino acid sequence shown in Seq.ID.No.:2 mBCNG-1 (FIG. 8A), Seq.ID.No.:6 for mBCNG-2 (FIG. 8A), Seq.ID.No.:10 for mBCNG-3 (FIG. 8A), Seq.ID.No.12 for mBCNG-4 (FIG. 8A), Seq.ID.No.:4 for hBCNG-1 (FIG. 8A), Seq.ID.No.:8 for hBCNG-2 (FIG. 8A) or a portion thereof.  
     
     
         22 . A nucleic acid probe capable of specifically hybridizing with a nucleic acid encoding a BCNG protein or BCNG-related protein.  
     
     
         23 . A nucleic acid probe capable of specifically hybridizing with the nucleic acid of  claim 1 .  
     
     
         24 . The nucleic acid probe of  claim 22 , wherein the probe is capable of specifically hybridizing with Seq.ID.No:13, Seq.ID.No:14, Seq.ID.No:15, Seq.ID.No:16, Seq.ID.No:17, Seq.ID.No:18, Seq.ID.No:19, Seq.ID.No:20, Seq.ID.No:21, Seq.ID.No:21, Seq.ID.No:22, Seq.ID.No:23, Seq.ID.No:24, Seq.ID.No:25, Seq.ID.No:26, Seq.ID.No:27, Seq.ID.No:28, Seq.ID.No:29, Seq.ID.No:30, Seq.ID.No:31, Seq.ID.No:32, Seq.ID.No:33, or Seq.ID.No:34.  
     
     
         25 . A method for identifying a nucleic acid in a sample which encodes a BCNG protein or a BCNG-related protein which comprises: 
 (a) contacting the sample with a nucleic acid probe capable of specifically hybridizing with nucleic acid encoding a BCNG protein or a BCNG-related protein under conditions permissive to the formation of a complex between the nucleic acid probe and the nucleic acid encoding the BCNG protein or the BCNG-related protein in the sample;    (b) determining the amount of complex formed in step (a); and    (c) comparing the amount of complex determined in step (b) with the amount of complex formed using an arbitrary sequence, a greater amount of complex formed With the BCNG-specific probe indicating the presence of a nucleic acid encoding a BCNG protein or a BCNG-related protein in the sample.    
     
     
         26 . The method of  claim 25 , step (a) further comprising amplifying the nucleic acid molecule encoding the BCNG protein or the BCNG-related protein.  
     
     
         27 . The method of  claim 26 , wherein the amplification comprises contacting the nucleic acid molecule from the sample with at least one amplification primer capable of specifically hybridizing to mBCNG-1 (Seq.ID.No.:1), mBCNG-2. (Seq.ID.No.:5)” mBCNG-3 (Seq.ID.No.:9), mBCNG-4 (Seq.ID.No.:11), hBCNG-1 (Seq.ID.No.:3) or hBCNG-2 (Seq.ID.No.:7) under conditions suitable for polymerase chain reaction.  
     
     
         28 . The method of  claim 26 , wherein the amplified nucleic acid molecule encoding the BCNG protein or the BCNG-related protein is detected by size fractionation.  
     
     
         29 . The method of  claim 25 , further comprising isolating the complex by size fractionation.  
     
     
         30 . The method of  claim 25 , wherein the nucleic acid probe is labeled with a detectable marker.  
     
     
         31 . The method of claim  3 d, wherein the detectable marker is a radiolabeled molecule, a fluorescent molecule, an enzyme, a ligand, or a magnetic bead.  
     
     
         32 . The method of  claim 25 , wherein the probe comprises Seq.ID.No:13, Seq.ID.No:14, Seq.ID.No:15, Seq.ID.No:16, Seq.ID.No:17, Seq.ID.No:18, Seq.ID.No:19, Seq.ID.No:20, Seq.ID.No:21, Seq.ID.No:21, Seq.ID.No:22, Seq.ID.No:23, Seq.ID.No:24, Seq.ID.No:25, Seq.ID.No:26, Seq.ID.No:27, Seq.ID.No:28, Seq.ID.No:29, Seq.ID.No:30, Seq.ID.No:31, Seq.ID.No:32, Seq.ID.No:33, or Seq.ID.No:34.  
     
     
         33 . The method of  claim 25 , wherein the nucleic acid probe is capable of specifically hybridizing to mBCNG-1 (Seq.ID.No.:1), mBCNG-2 (Seq.ID.No.:5), mBCNG-3 (Seq.ID.No.:9), mBCNG-4 (Seq.ID.No.:11), hBCNG-1 (Seq.ID.No.:3) or hBCNG-2 (Seq.ID.No.:7).  
     
     
         34 . An isolated nucleic acid, previously unknown, identified by the method of  claim 25 .  
     
     
         35 . A method for testing whether a compound affects the expression of a BCNG protein or a BCNG-related protein which comprises: 
 (a) contacting a sample which expresses a BCNG protein or a BCNG-related protein with a compound;    (b) determining the amount of expression of BCNG protein or BCNG-related protein in the sample; and    (c) comparing the amount of BCNG protein or BCNG-related protein expression determined in step (b) with the amount determined in the absence of the compound.    
     
     
         36 . A method for identifying a compound capable of interacting with a BCNG protein or a BCNG-related protein which comprises: 
 (a) contacting a sample which expresses a BCNG protein or a BCNG-related protein with a compound under conditions permissive to formation of a complex between the compound and the BCNG protein or the BCNG-related protein;    (b) determining the amount of complex formed between the compound and the BCNG protein or the BCNG-related protein;    (c) comparing the amount of complex formed in step (b) with the amount formed in the absence of the compound, a greater amount of complex formed in the presence of the compound indicating the presence of a compound capable of interacting with a BCNG protein or a BCNG-related protein.    
     
     
         37 . A method for identifing a compound capable of modulating BCNG protein or BCNG-related protein activity which comprises: 
 (a) contacting a sample which expresses a BCNG protein or a BCNG-related protein with a compound;    (b) determining the amount of activity of the BCNG protein or BCNG-related protein in the sample; and    (c) comparing the amount of activity of the BCNG protein or the BCNG-related protein determined in step (b) with the amount determined in the absence of the compound, an increase or decrease in activity indicating the presence of a compound capable of modulating the activity of the BCNG protein or the BCNG-related protein.    
     
     
         38 . The method of  claim 37 , step (a) comprising first introducing the nucleic acid encoding a BCNG protein or a BCNG-related protein into an expression system and causing the expression system to express the nucleic acid under conditions whereby a BCNG protein or a BCNG-related protein is produced.  
     
     
         39 . The method of  claim 37 , wherein step (b) comprises measuring the channel electrical current or intracellular calcium level in the presence of the compound.  
     
     
         40 . The method of  claim 38 , wherein the expression system comprises a cultured host cell.  
     
     
         41 . A compound, previously unknown, identified by the method of  claim 35 ,  36 , or  37 .  
     
     
         42 . The method of  claim 25 ,  35 ,  36 , or  37 , wherein the BCNG protein or the BCNG-related protein comprises mBCNG-1 (Seq.ID.No.:2), mBCNG-2 (Seq.ID.No.:6), mBCNG-3 (Seq.ID.No.:10), mBCNG-4 (Seq.ID.No.:12), hBCNG-1 (Seq.ID.No.:4) or hBCNG-2 (Seq.ID.No.:8) or a portion thereof.  
     
     
         43 . The method of  claim 25 ,  35 ,  36 , or  37 , wherein the sample comprises a cell, cell lysate or cell-free translation.  
     
     
         44 . The method of  claim 43 , wherein the cell is a cardiac cell, a kidney cell, a hepatic cell, an airway epithelial cell, a muscle cell, a neuronal cell, a glial cell, a microglial cell, an endothelial cell, a mononuclear cell, a tumor cell, a mammalian cell, an insect cell, or a Xenopus oocyte.  
     
     
         45 . A compound, previously unknown, identified by the method of  claim 35 ,  36 , or  37 .  
     
     
         46 . The method of  claim 35 ,  36  or  37 , wherein the compound is a peptide, a peptidomimetic, a nucleic acid, a polymer, or a small molecule.  
     
     
         47 . The method of  claim 35 ,  36  or  37 , wherein the compound is bound to a solid support.  
     
     
         48 . The method of  claim 25 ,  35 ,  36 , or  37 , wherein the BCNG protein or the BCNG-related protein is ion channel protein or an ion channel subunit protein.  
     
     
         49 . The method of  claim 35 ,  36 , or  37 , wherein the compound is an agonist or antagonist of ion channel activity  
     
     
         50 . The method of  claim 37  wherein the modulation is increased ion flow rate or decreased ion flow rate.  
     
     
         51 . The method of  claim 37  wherein the modulation is increased ion permissivity or decreased ion permissivity  
     
     
         52 . A method of modulating BCNG or BCNG-related protein activity in a sample, comprising contacting the sample with the compound of  claim 41 .  
     
     
         53 . A method of treating a condition in a subject which comprises administering to the subject an amount of the compound of  claim 41 , effective to treat the condition.  
     
     
         54 . A pharmaceutical composition which comprises the compounds of  claim 41  and a pharmaceutically acceptable carrier.  
     
     
         55 . The pharmaceutical composition of  claim 54 , wherein the carrier is a diluent, an aerosol, a topical carrier, an aqueous solution, a nonaqueous solution or a solid carrier.  
     
     
         56 . A method for treating a condition in a subject which comprises administering to the subject an amount of the pharmaceutical composition of  claim 54 , effective to treat the condition in the subject.  
     
     
         57 . The method of  claim 53  or  56 , wherein the condition is a neurological, renal, pulmonary, hepatic, or cardiovascular condition.  
     
     
         58 . The method of  claim 57 , wherein the condition is epilepsy, Alzheimer's Disease, Parkinson's Disease, long QT syndrome, sick sinus syndrome, age-related memory loss, cystic fibrosis, sudden death syndrome or a pacemaker rhythm dysfunction.  
     
     
         59 . The method of  claim 53  or  56 , wherein the subject is a human.  
     
     
         60 . An antibody which binds specifically to the protein of  claim 14  or  15 .  
     
     
         61 . A cell capable of producing the antibody of  claim 60 .  
     
     
         62 . A method of identifying the protein of  claim 14  or in a sample comprising: 
 a) contacting the sample with the antibody of  claim 58  under conditions permissive to the formation of a complex between the antibody and the protein;  
 b) determining the amount of complex formed; and  
 c) comparing the amount of complex formed in step (b) with the amount of complex formed in the absence of the antibody, the presence of an increased amount of complex formed in the presence of the antibody indicating identification of the protein in the sample.