Isolated nucleic acid molecules encoding a novel human signal transducing kinase-mapkap-2; encoded proteins, cells transformed therewith and uses thereof

An isolated nucleic acid molecule, comprising a coding region that encodes a splice variant of a MAPKAP-2 kinase, wherein the MAPKAP-2 kinase is encoded by a sequence of nucleotides as set forth in SEQ ID NO:1.

An isolated nucleic acid molecule that encodes a MAPKAP-2 kinase having an amino acid sequence as set forth in SEQ ID NO:2.

A substantially pure human MAPKAP-2 kinase encoded by a nucleotide sequence that is a splice variant of a isolated nucleic acid molecule that encodes a MAPKAP-2 kinase comprising the amino acid sequence set forth in SEQ ID NO:2.

A substantially pure human MAPKAP-2 kinase encoded by a nucleotide sequence as set forth in SEQ ID NO:1.

A substantially pure human MAPKAP-2 kinase encoded by a nucleic acid molecule comprising a sequence of nucleotides that hybridizes under stringent wash conditions to the complement of the sequence of nucleotides set forth in SEQ ID NO:1.

A substantially pure human MAPKAP-2 kinase comprising a sequence of amino acids as set forth in SEQ ID NO:

• 2.

A method for determining regression, progression or onset of a disease state manifested by a dysfunctional signal transducing MAPKAP-2 kinase, the method comprising the steps of (i) contacting a cDNA or mRNA containing sample from a subject suspected of suffering from said disease state with a nucleic acid hybridization probe comprising a sequence of nucleotides as set forth in SEQ ID NO:

• 1 under conditions favoring binding of the hybridization probe to the cDNA or mRNA to form a complex therebetween, and (ii) detecting said complex as an indication that said subject is at risk of developing said disease state.

A method for determining regression, progression or onset of a pathological disorder characterized by a dysfunctional signal transducing MAPKAP-2 kinase comprising:

• contacting a sample from a patient with the disorder with a detectable probe that is specific for the gene product of the isolated nucleic acid molecule having a sequence of nucleotides as set forth in SEQ ID NO:1 under conditions favoring formation of a probe/gene product complex, the presence of which is indicative of the regression, progression or onset of the pathological disorder in the patient.

A method of screening test compounds for use as inflammation inhibitors, comprising the steps of:

• a) contacting a test compound with a MAPKAP-2 kinase encoded by a polynucleotide comprising a nucleotide sequence selected from the group consisting of SEQ ID NO:1 and a sequence of nucleotides that encodes a human MAPKAP-2 kinase that hybridizes under conditions of high stringency to the complement of the sequence of nucleotides set forth in SEQ ID NO:1;
• and
• b) testing the contacted MAPKAP-2 kinase protein for its ability to bind to or phosphorylate Hsp-27, wherein a test compound which inhibits phosphorylation of Hsp-27 by the MAPKAP-2 kinase or which inhibits the binding of the MAPKAP-2 kinase protein to the Hsp-27 is a candidate drug for treatment of inflammation.

An isolated nucleic acid molecule comprising a nucleotide sequence encoding a polypeptide that has at least 80% identity to the amino acid sequence set forth SEQ ID NO:2 over the entire length of SEQ ID NO:2, wherein 80% identity defines the amino acid alterations allowed for SEQ ID NO:2 which are determined by the equation and is calculated by the formula

• Na=Xa−(Xa Y),
• wherein Na is the maximum number of amino acid alterations, Xa is the total number of amino acids in SEQ ID NO:2, and Y has a value of 0.80, wherein any non-integer product of Xa and Y is rounded down to the nearest integer prior to subtracting such product from Xa.

A method for detecting a binding partner for a MAPKAP-2 kinase in a sample suspected of containing the binding partner, comprising:

• (i) contacting the sample with the MAPKAP-2 of SEQ ID NO:2 under conditions favoring binding of the MAPKAp-2 kinase to the binding partner;
• (ii) determining presence of the binding partner in the sample by detecting binding of the MAPKAP-2 kinase to the binding partner.

A method for identifying a compound which modulates the binding or kinase activity of a naturally occurring allelic variant of a polypeptide consisting of the amino acid sequence of SEQ ID NO:2, wherein the allelic variant is encoded by a nucleic acid molecule which hybridizes to the complement of a nucleic acid molecule consisting of SEQ ID NO:1 in 6×SSC at 45° C., followed by one or more washes in 0.2.times.SSC, 0.1% SDS at 50-65° C., the method comprising:

• a) contacting a cell expressing the allelic variant with a test compound under conditions suitable for modulation of the binding or kinase activity of the allelic variant;
• and
• b) detecting modulation of the binding or kinase activity of the allelic variant by the test compound.

Isolated polypeptide comprising an amino acid sequence provided in SEQ ID NO:2, or a variant thereof that is at least 80% identical to SEQ ID NO:2 and that differs from SEQ ID NO:2 only in one or more amino acid substitutions, additions of terminal amino acid residues and/or deletions of terminal amino acid residues, wherein the ability of the variant to phosphorylate Hsp-27 is not substantially diminished.

A method of treating a subject having a disorder characterized by aberrant MAPKAP-2 kinase or nucleic acid expression or activity comprising administering an agent which is a MAPKAP-2 modulator to the subject.

A method of phosphorylating a serine-containing substrate which comprises:

• a) incubating the substrate with an effective concentration of ATP and an enzyme having at least 84% homology to SEQ ID NO:1, wherein said enzyme phosphorylates Hsp-27;
• and
• b) measuring the amount of phosphorylation of the substrate.

A kit useful for the detection of mitogen-activating protein kinase activating protein kinase 2 (MAPKAP-2), said kit comprising a buffer and a labeled antibody which specifically binds to a MAPKAP-2 kinase having serine, threonine, and tyrosine kinase activity, wherein the sample to be tested is mixed with the buffer and the antibody.

A kit useful for the detection of mitogen-activating protein kinase activating protein kinase 2 (MAPKAP-2) encoding nucleic acid, said kit comprising a buffer and nucleic acid molecule comprising at least about 20 nucleotides capable of hybridizing to a nucleic acid sequence encoding MAPKAP-2 or a complement thereof under stringent hybridization conditions, and instructions for use thereof.

An isolated nucleic acid molecule, comprising a coding region that encodes a splice variant of a MAPKAP-2 kinase, wherein the MAPKAP-2 kinase is encoded by a sequence of nucleotides as set forth in SEQ ID NO:

• 3.

An isolated nucleic acid molecule that encodes a MAPKAP-2 kinase having an amino acid sequence as set forth in SEQ ID NO:4.

A substantially pure MAPKAP-2 kinase encoded by a nucleotide sequence that is a splice variant of a isolated nucleic acid molecule that encodes a MAPKAP-2 kinase comprising the amino acid sequence set forth in SEQ ID NO:4.

An isolated nucleic acid molecule comprising a nucleotide sequence having at least 80% identity to a nucleotide sequence encoding a polypeptide comprising the amino acid sequence set forth SEQ ID NO:2, which may include up to Na nucleic acid alterations over the entire length of SEQ ID NO:1, wherein Na represents the maximum number of such alterations and is calculated by the formula

• Na=Xa−(Xa Y),
• In which Xa is the total number of nucleotides in SEQ D NO:1, and Y has a value of 0.80, wherein any non-integer product of Xa and Y is rounded down to the nearest integer prior to subtracting such product from Xa, wherein said polypeptide has MAPKAp-2 activity.