Human Ependymin
The present invention relates to a novel human Ependymin protein which is a member of the ependymin family. In particular, isolated nucleic acid molecules are provided encoding the human Ependymin protein. Ependymin polypeptides are also provided as are vectors, host cells and recombinant methods for producing the same. The invention further relates to screening methods for identifying agonists and antagonists of Ependymin activity. Also provided are diagnostic methods for detecting nervous system-related disorders and therapeutic methods for treating nervous system-related disorders.
1 . An isolated nucleic acid molecule nucleic acid molecule comprising a polynucleotide having a nucleotide sequence at least 95% identical to a sequence selected from the group consisting of:
(a) a nucleotide sequence encoding the Ependymin polypeptide having the complete amino acid sequence in SEQ ID NO:2 (i.e., positions −37 to 187 of SEQ ID NO:2);
(b) a nucleotide sequence encoding the Ependymin polypeptide having the complete amino acid sequence in SEQ ID NO:2 excepting the N-terminal methionine (i.e., positions −36 to 187 of SEQ ID NO:2);
(c) a nucleotide sequence encoding the predicted mature Ependymin polypeptide having the amino acid sequence at positions 1 to 187 in SEQ ID NO:2;
(d) a nucleotide sequence encoding the Ependymin polypeptide having the complete amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit No. 209464;
(e) a nucleotide sequence encoding the Ependymin polypeptide having the complete amino acid sequence excepting the N-terminal methionine encoded by the cDNA clone contained in ATCC™ Deposit No. 209464;
(f) a nucleotide sequence encoding the mature Ependymin polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit No. 209464; and
(g) a nucleotide sequence complementary to any of the nucleotide sequences in (a), (b), (c), (d), (e) or (f), above.
2 . The nucleic acid molecule of claim 1 wherein said polynucleotide has the complete nucleotide sequence in FIGS. 1A, 1B , and 1 C (SEQ ID NO:1).
3 . The nucleic acid molecule of claim 1 wherein said polynucleotide has the nucleotide sequence in FIGS. 1A, 1B , and 1 C (SEQ ID NO:1) encoding the mature Ependymin polypeptide having the amino acid sequence from about 1 to about 187 in SEQ ID NO:2.
4 . The nucleic acid molecule of claim 1 wherein said polynucleotide has the complete nucleotide sequence of the cDNA clone contained in ATCC™ Deposit No. 209464.
5 . The nucleic acid molecule of claim 1 wherein said polynucleotide has the nucleotide sequence encoding the Ependymin polypeptide having the complete amino acid sequence excepting the N-terminal methionine encoded by the cDNA clone contained in ATCC™ Deposit No. 209464.
6 . The nucleic acid molecule of claim 1 wherein said polynucleotide has the nucleotide sequence encoding the mature Ependymin polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit No. 209464.
7 . An isolated nucleic acid molecule comprising a polynucleotide which hybridizes under stringent hybridization conditions to a polynucleotide having a nucleotide sequence identical to a nucleotide sequence in (a), (b), (c), (d), (e), (f) or (g) of claim 1 wherein said polynucleotide which hybridizes does not hybridize under stringent hybridization conditions to a polynucleotide having a nucleotide sequence consisting of only A residues or of only T residues.
8 . An isolated nucleic acid molecule comprising a polynucleotide which encodes the amino acid sequence of an epitope-bearing portion of a Ependymin polypeptide having an amino acid sequence in (a), (b), (c), (d), (e) or (f) of claim 1 .
9 . The isolated nucleic acid molecule of claim 10 , which encodes an epitope-bearing portion of a Ependymin polypeptide wherein the amino acid sequence of said portion is selected from the group of sequences in SEQ ID NO:2 consisting of: about Ala-1 to about Gln-9; about Pro-8 to about Val-16; about Gln-19 to about Arg-27; about Ile-69 to about Ser-77; about Asp-86 to about Glu-107; about Glu-113 to about Tyr-123; about Thr-131 to about Gln-139; about Leu-159 to about Phe-167; and about Leu-178 to about Ser-186.
10 . A method for making a recombinant vector comprising inserting an isolated nucleic acid molecule of claim 1 into a vector.
11 . A recombinant vector produced by the method of claim 10 .
12 . A method of making a recombinant host cell comprising introducing the recombinant vector of claim 11 into a host cell.
13 . A recombinant host cell produced by the method of claim 12 .
14 . A recombinant method for producing an Ependymin polypeptide, comprising culturing the recombinant host cell of claim 13 under conditions such that said polypeptide is expressed and recovering said polypeptide.
15 . An isolated Ependymin polypeptide comprising an amino acid sequence at least 95% identical to a sequence selected from the group consisting of:
(a) the amino acid sequence of the full-length Ependymin polypeptide having the complete amino acid sequence shown in SEQ ID NO:2 (i.e., positions −37 to 187 of SEQ ID NO:2);
(b) the amino acid sequence of the full-length Ependymin polypeptide having the complete amino acid sequence shown in SEQ ID NO:2 excepting the N-terminal methionine (i.e., positions −36 to 187 of SEQ ID NO:2);
(c) the amino acid sequence of the predicted mature Ependymin polypeptide having the amino acid sequence at positions 1 to 187 in SEQ ID NO:2;
(d) the complete amino acid sequence encoded by the cDNA clone contained in the ATCC™ Deposit No. 209464;
(e) the complete amino acid sequence excepting the N-terminal methionine encoded by the cDNA clone contained in the ATCC™ Deposit No. 209464; and
(f) the complete amino acid sequence of the predicted mature Ependymin polypeptide encoded by the cDNA clone contained in the ATCC™ Deposit No. 209464.
16 . An isolated antibody that binds specifically to a Ependymin polypeptide of claim 15 .
17 . An isolated nucleic acid molecule comprising a polynucleotide having a sequence at least 95% identical to a sequence selected from the group consisting of:
(a) the nucleotide sequence of SEQ ID NO:15;
(b) the nucleotide sequence of SEQ ID NO:16;
(c) the nucleotide sequence of SEQ ID NO:17;
(d) the nucleotide sequence of SEQ ID NO:18;
(e) the nucleotide sequence of SEQ ID NO:19;
(f) the nucleotide sequence of SEQ ID NO:20;
(g) the nucleotide sequence of a portion of the sequence shown in FIGS. 1A, 1B , and 1 C (SEQ ID NO:1) wherein said portion comprises at least 50 contiguous nucleotides from nucleotide 1 to nucleotide 2505; and
(h) a nucleotide sequence complementary to any of the nucleotide sequences in (a), (b), (c), (d), (e), (f) or (g), above.
18 . A method for the treatment of a patient having need of human ependymin polypeptide comprising:
(a) isolating the polypeptide of claim 1 , and
(b) administering to the patient a therapeutically effective amount of said polypeptide.
19 . A method for identifying compounds which bind to and inhibit activation of the polypeptide of claim 1 comprising:
(a) contacting a cell expressing on the surface thereof a receptor for the polypeptide, said receptor being associated with a second component capable of providing a detectable signal in response to the binding of a compound to said receptor, with an analytically detectable human cytokine polypeptide and a compound under conditions to permit binding to the receptor; and
(b) determining whether the compound binds to and inhibits the receptor by detecting the absence of a signal generated from the interaction of the human ependymin polypeptide with the receptor.
20 . A process for diagnosing a disease or a susceptibility to a disease related to an under-expression of the polypeptide of claim 1 comprising:
(a) isolating a sample of the nucleic acid sequence encoding said polypeptide, and
(b) determining a mutation in a nucleic acid sequence encoding said polypeptide.