Human calcium channel compositions and methods
AssigneeMerck & Co., Inc.
InventorsMichael M. Harpold, Steven B. Ellis, Mark E. Williams, Ann F. McCue
Life of this patent
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Abstract
Isolated DNA encoding each of human calcium channel α1-, α2-, β- and γ-subunits, including subunits that arise as splice variants of primary transcripts, is provided. Cells and vectors containing the DNA and methods for identifying compounds that modulate the activity of human calcium channels are also provided.
Claims24
A eukaryotic cell, comprising heterologous DNA that encodes an α1 subunit of a human calcium channel, and heterologous DNA that encodes a full-length β-subunit of a human calcium channel encoded by a sequence of nucleotides selected from the group consisting of:
- (a) a sequence of nucleotides that encodes a human β3-subunit and that comprises the sequence of nucleotides set forth in SEQ ID No. 20;
- (b) a sequence of nucleotides that encodes a β3-subunit, wherein:
- a DNA molecule having said sequence hybridizes under conditions of high stringency to DNA that is fully complementary to an mRNA transcript native to a human cell;
- and
- the mRNA transcript encodes a β3-subunit comprising the coding portion of the sequence of nucleotides set forth in SEQ ID No. 19 or 20;
- (c) a sequence of nucleotides that encodes a β3-subunit that comprises the sequence of amino acids encoded by SEQ ID No. 20;
- (d) a sequence of nucleotides that encodes the human β4-subunit having SEQ ID No. 28;
- (e) a sequence of nucleotides that encodes a β4-subunit, wherein:
- a DNA molecule having said sequence hybridizes under conditions of high stringency to DNA that is fully complementary to an mRNA transcript native to a human cell;
- and
- the mRNA transcript encodes a β4-subunit comprising the coding portion of the sequence of nucleotides set forth in SEQ ID No. 28;
- (f) a sequence of nucleotides that encodes a human β3-subunit and that comprises the sequence of nucleotides set forth in SEQ ID No. 19;
- and
- (g) a sequence of nucleotides that encodes a β3-subunit that comprises the sequence of amino acids encoded by SEQ ID No. 19;
- wherein the cell has a functional heterologous calcium channel that contains at least one subunit encoded by the heterologous DNA that encodes a full-length β-subunit of a human calcium channel.
A eukaryotic cell with a functional, heterologous calcium channel, produced by a process comprising:
- introducing into the cell heterologous nucleic acid that encodes at least one subunit of a calcium channel, wherein:
- the heterologous nucleic acid comprises a sequence of nucleotides that encodes a full-length β3-subunit of a human calcium channel, and is fully complementary to mRNA that encodes the β3-subunit of a human calcium channel encoded by SEQ ID No. 19 or 20 and is native to a human cell;
- the cell further comprises nucleic acid that encodes an α1 subunit of a human calcium channel;
- the heterologous calcium channel contains at least one subunit encoded by the heterologous nucleic acid.
A eukaryotic cell, comprising heterologous nucleic acid that encodes an α1 subunit of a human calcium channel, an α2 subunit of a human calcium channel, a γ subunit of a calcium channel and heterologous nucleic acid that encodes a β3 subunit of a human calcium channel, wherein the heterologous nucleic acid that encodes a β3 subunit of a human calcium comprises:
- (a) a sequence of nucleotides that encodes a human β3-subunit and that comprises the sequence of nucleotides set forth in SEQ ID No. 20;
- (b) a sequence of nucleotides that encodes a human β3-subunit that comprises the sequence of amino acids encoded by SEQ ID No. 20;
- (c) a sequence of nucleotides that encodes a human β3-subunit and that comprises the sequence of nucleotides set forth in SEQ ID No. 19;
- or
- (b) a sequence of nucleotides that encodes a human β3-subunit that comprises the sequence of amino acids encoded by SEQ ID No. 19.
An isolated DNA molecule encoding a β3 subunit where the isolated DNA molecule comprises the β3-encoding sequence of nucleotides contained in a plasmid having all of the identifying characteristics of the plasmid harbored in the Escherichia coli(E. coli) host cells having ATCC Accession No. 69048.
An isolated RNA molecule degenerate with a DNA molecule which comprises the sequence of nucleotides set forth in SEQ ID No. 19 or 20.
Dependencies are inferred from the claim text.