PARG, a GTPase activating protein which interacts with PTPL1
OwnerJan Saras, Petra Franzen +4inventors
InventorsJan Saras, Petra Franzen, Pontus Aspenstrom, Ulf Hellman, Leonel Jorge Gonez, Carl-Henrik Heldin
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Abstract
The invention describes nucleic acids encoding the PARG protein, including fragments and biologically functional variants thereof. Also included are polypeptides and fragments thereof encoded by such nucleic acids, and antibodies relating thereto. Methods and products for using such nucleic acids and polypeptides also are provided.
Claims53
An isolated nucleic acid molecule
- (a) which hybridizes under stringent conditions to a molecule consisting of the nucleic acid sequence of SEQ ID NO:1 and which codes for a GTPase-activating polypeptide,
- (b) nucleic acid molecules that differ from the nucleic acid molecules of (a) in codon sequence due to the degeneracy of the genetic code, and
- (c) complements of (a) and (b).
An isolated nucleic acid molecule selected from the group consisting of (a) a unique fragment of nucleotides 184-3966 of SEQ ID NO:1 between 12 and 3781 nucleotides in length and (b) complements of “(a)”, provided that the nucleic acid molecule excludes molecules consisting solely of nucleotide sequences selected from the group consisting of accession numbers T32345, Z28937, Z28520, AA431926, AA326126, AA342471, AA716829, L49573, Z43348, AA303722, T32495, AA330162, Z25350, AA794256, T32506, T32263, F06673, AA462548, X85558, R14952, AA870705, AA120493, AA415591, AA131400, C76597, C76601, AA870475, AA234871, C77518, and AA672012.
An isolated nucleic acid molecule, comprising
- (a) a molecule having a sequence selected from the group consisting of SEQ ID NO:6, SEQ ID NO:8 and SEQ ID NO:10;
- (b) nucleic acid molecules that differ from the nucleic acid molecules of (a) in codon sequence due to the degeneracy of the genetic code, and
- (c) complements of (a) and (b).
An isolated polypeptide, comprising a polypeptide having the sequence of amino acids 613-652 of SEQ ID NO:2, wherein the isolated polypeptide is a cysteine-rich domain.
An isolated polypeptide, comprising a polypeptide having the sequence of amino acid 193-509 of SEQ ID NO:2, wherein the isolated polypeptide is a ZPH domain polypeptide.
An isolated polypeptide comprising a polypeptide selected from the group consisting of a polypeptide having the sequence of SEQ ID NO:7, a polypeptide having the sequence of SEQ ID NO:9, and a polypeptide having the sequence of SEQ ID NO:11.
A method for reducing the proliferation of a cancer cell, the proliferation of which is increased by Rho family protein signal transduction, comprising
- administering to the cancer cell an amount of a polypeptide comprising a polypeptide encoded by the nucleic acid of SEQ ID NO:1 effective to reduce proliferation of the cancer cell.
A method of increasing Rho family protein signal transduction in a cell comprising
- administering to the cell an amount of a dominant-negative variant of the polypeptide of SEQ ID NO:2 effective to increase the Rho family protein signal transduction in the cell.
A method of reducing binding of a protein which includes a PDZ4 domain to a protein which includes a PDZ4 domain binding site in a mammalian cell, comprising
- contacting the mammalian cell with an agent which binds to the PDZ4 domain for a time effective to reduce binding of the protein which includes a PDZ4 domain to the protein which includes a PDZ4 domain binding site.
42 The method of claim 41, wherein the antibody is a monoclonal antibody.
A method of modulating mast cell secretion in a subject, comprising:
- administering to the subject in need of such treatment an amount of a modulator of PARG GTPase activating activity effective to modulate mast cell secretion.
A method for identifying lead compounds for a pharmacological agent useful in the diagnosis or treatment of disease associated with PARG GTPase activating activity, comprising
- forming a mixture comprising a PARG polypeptide or unique fragment thereof containing a PARG GTPase activating domain, a protein containing a GTPase specifically activatable by the PARG GTPase activating domain, and a candidate pharmacological agent,
- incubating the mixture under conditions which, in the absence of the candidate pharmacological agent, permit a first amount of specific activation of the GTPase by the PARG GTPase activating domain, and
- detecting a test amount of the specific activation of the GTPase by the PARG GTPase activating domain, wherein reduction of the test amount of specific activation relative to the first amount of specific activation indicates that the candidate pharmacological agent is a lead compound for a pharmacological agent which disrupts the GTPase activation activity of PARG, and wherein increase of the test amount of specific activation relative to the first amount of specific activation indicates that the candidate pharmacological agent is a lead compound for a pharmacological agent which enhances the GTPase activation activity of PARG.
A method for identifying lead compounds for a pharmacological agent useful in the diagnosis or treatment of disease associated with PARG binding to a protein containing a PDZ4 domain, comprising
- forming a mixture comprising a PARG polypeptide or unique fragment thereof containing a PDZ4 domain binding site, a protein containing a PDZ4 domain which selectively binds the PARG PDZ4 domain binding site, and a candidate pharmacological agent,
- incubating the mixture under conditions which, in the absence of the candidate pharmacological agent, permit a first amount of selective binding of the protein containing a PDZ4 domain by the PDZ4 domain binding site, and
- detecting a test amount of selective binding of the protein containing a PDZ4 domain by the PDZ4 domain binding site, wherein reduction of the test amount of selective binding relative to the first amount of selective binding indicates that the candidate pharmacological agent is a lead compound for a pharmacological agent which disrupts selective binding of a protein containing a PDZ4 domain by a PARG polypeptide containing a PDZ4 domain binding site and wherein increase of the test amount of selective binding relative to the first amount of selective binding indicates that the candidate pharmacological agent is a lead compound for a pharmacological agent which enhances selective binding of a protein containing a PDZ4 domain by a PARG polypeptide containing a PDZ4 domain binding site.
Dependencies are inferred from the claim text.