Regulation of novel human asparagine-hydroxylases
Reagents that regulate human asparagine-hydroxylase and reagents which bind to human asparagine-hydroxylase gene products can play a role in preventing, ameliorating, or correcting dysfunctions or diseases including, but not limited to cardiovascular disorders, anaemia, cancer, inflammatory diseases, fibrotic disorders, and CNS disorders.
1 . An isolated polynucleotide being selected from the group consisting of:
a) a polynucleotide encoding an asparagine-hydroxylase polypeptide comprising an amino acid sequence selected from the group constisting of:
i) amino acid sequences which are at least about 36% identical to the amino acid sequence shown in SEQ ID NO: 2; and
ii) the amino acid sequence shown in SEQ ID NO: 2;
iii) amino acid sequences which are at least about 38% identical to the amino acid sequence shown in SEQ ID NO: 4; and
iv) the amino acid sequence shown in SEQ ID NO: 4;
v) amino acid sequences which are at least about 28% identical to the amino acid sequence shown in SEQ ID NO: 6; and
vi) the amino acid sequence shown in SEQ ID NO: 6.
b) a polynucleotide comprising the sequence of SEQ ID NO: 1, 3 or 5;
c) a polynucleotide which hybridizes under stringent conditions to a polynucleotide specified in (a) and (b) and encodes an asparagine-hydroxylase;
d) a polynucleotide the nucleic acid sequence of which deviates from the nucleic acid sequences specified in (a) to (c) due to the degeneration of the genetic code and encodes an asparagine-hydroxylase; and
e) a polynucleotide, which represents a fragment, derivative or allelic variation of a nucleic acid sequence specified in (a) to (d) and encodes an asparagine-hydroxylase.
2 . An expression vector containing any polynucleotide sequence of claim 1 .
3 . A host cell containing the expression vector of claim 2 .
4 . A substantially purified asparagine-hydroxylase polypeptide encoded by a polynucleotide of claim 1 .
5 . A method for producing an asparagine-hydroxylase polypeptide, wherein the method comprises the steps of:
a) culturing the host cell of claim 3 under conditions suitable for the expression of the polypeptide; and
b) recovering the asparagine-hydroxylase polypeptide from the host cell culture.
6 . A method for the detection of a polynucleotide encoding asparagine-hydroxylase polypeptide in a biological sample comprising the following steps:
a) hybridizing any polynucleotide of claim 1 to nucleic acid material of a biological sample, thereby forming a hybridization complex; and
b) detecting said hybridization complex.
7 . The method of claim 6 , wherein before hybridization, the nucleic acid material of the biological sample is amplified.
8 . A method for the detection of a polynucleotide of claim 1 or an asparagine-hydroxylase polypeptide of claim 4 comprising the steps of:
a) contacting a biological sample with a reagent which specifically interacts with the polynucleotide or the asparagine-hydroxylase poly-peptide; and
b) detecting the interaction.
9 . A diagnostic kit for conducting the method of one of the claims 6 to 8 .
10 . A method of screening for agents which decrease the activity of an asparagine-hydroxylase, comprising the steps of:
contacting a test compound with any asparagine-hydroxylase polypeptide encoded by any polynucleotide of claim 1;
detecting binding of the test compound to the asparagine-hydroxylase polypeptide, wherein a test compound which binds to the polypeptide is identified as a potential therapeutic agent for decreasing the activity of an asparagine-hydroxylase.
11 . A method of screening for agents which regulate the activity of an asparagine-hydroxylase, comprising the steps of:
a) contacting a test compound with an asparagine-hydroxylase poly-peptide encoded by any of the polynucleotides of claim 1; and
b) detecting the asparagine-hydroxylase activity of the polypeptide, wherein a test compound which increases the asparagine-hydroxylase activity is identified as a potential therapeutic agent for increasing the activity of the asparagine-hydroxylase and wherein a test compound which decreases the asparagine-hydroxylase activity of the polypeptide is identified as a potential therapeutic agent for decreasing the activity of the asparagine-hydroxylase.
12 . A method of screening for agents which decrease the activity of an asparagine-hydroxylase, comprising the steps of:
a) contacting a test compound with any polynucleotide of claim 1; and
b) detecting binding of the test compound to the polynucleotide, wherein a test compound which binds to the polynucleotide is identified as a potential therapeutic agent for decreasing the activity of the asparagine-hydroxylase
13 . A method of reducing the activity of an asparagine-hydroxylase comprising the step of:
contacting a cell with a reagent which specifically binds to any polynucleotide of claim 1 or an asparagine-hydroxylase polypeptide of claim 4 , whereby the activity of the asparagine-hydroxylase is reduced.
14 . A reagent that modulates the activity of an asparagine-hydroxylase poly-peptide or polynucleotide, wherein said reagent is identified by the method of any of the claims 10 to 12 .
15 . A pharmaceutical composition, comprising:
the expression vector of claim 2 or the reagent of claim 14 , and a pharmaceutically acceptable carrier.
16 . Use of the expression vector of claim 2 or the reagent of claim 14 in the preparation of a medicament for modulating the activity of an asparagine-hydroxylase in a disease.
17 . Use of claim 16 , wherein the disease is a cardiovascular disorder, anaemia, cancer, an inflammatory disease, a fibrotic disorder or a CNS disorder.