Optical Disc Apparatus

Abstract

Disclosed are human mitogen-activated (MAP) kinase kinase isoforms (MKKs). MKKs mediate unique signal transduction pathways that activate human MAP kinases p38 and JNK, which result in activation of other factors, including activating transcription factor-2 (ATF2) and c-Jun. The pathways are activated by a number of factors, including cytokines and environmental stress. Methods are provided for identifying reagents that modulate MKK function or activity and for the use of such reagents in the treatment of MKK-mediated disorders.

1 .- 37 . (canceled) 
     
     
         38 . A method of measuring the activity of a mitogen-activated protein kinase kinase (MKK) in a biological test sample, said method comprising:
 a) incubating said test sample with an MKK substrate for a substantially pure human mitogen-activated protein kinase kinase (MKK) polypeptide having serine, threonine, and tyrosine kinase activity, and phosphorylating human mitogen-activated protein (MAP) kinase p38, and labeled phosphate under conditions sufficient to allow phosphorylation of said substrate, and   b) determining the rate of incorporation of labeled phosphate into said substrate, wherein said rate of incorporation is a measure of MKK activity.   
     
     
         39 . A method of  claim 38  wherein said MKK substrate is selected from the group consisting of p38 and JNK MAP kinases, activating transcription factor-2 (ATF2), ATFa, cAMP response element binding protein (CRE-BPa), and c-Jun. 
     
     
         40 . A method of  claim 38  wherein said biological test sample is fluid, cells, or tissue obtained from a mammal. 
     
     
         41 . A method for measuring the level of expression of MKK in a test sample, comprising the steps of:
 a) isolating polyadenylated RNA from the test sample;   b) incubating polyadenylated RNA with a polynucleotide probe specific for a substantially pure human mitogen-activated protein kinase kinase (MKK) polypeptide having serine, threonine, and tyrosine kinase activity, and phosphorylating human mitogen-activated protein (MAP) kinase p38;   c) determining the amount of said probe hybridized said polyadenylated RNA, wherein the level of expression of MKK is directly related to the amount of MKK probe hybridized to said RNA.   
     
     
         42 . A method of treating an MKK-mediated disorder in a patient, the method comprising administering to the patient a therapeutically effective amount of a reagent that modulates MKK activity. 
     
     
         43 . The method of  claim 42 , wherein the MKK-mediated disorder is selected from the group consisting of ischemic heart disease, kidney failure, oxidative liver damage, respiratory distress syndrome, burns from heat, radiation burns, septic shock, rheumatoid arthritis, autoimmune disorders, and inflammatory diseases. 
     
     
         44 . The method of  claim 43 , wherein the radiation burn is caused by exposure to UV, X-rays, γ particles, or β particles. 
     
     
         45 . The method of  claim 42 , wherein the MKK-mediated disorder is selected from the group consisting of a proliferative disorder, psoriasis, acquired immune deficiency syndrome, and malignancy. 
     
     
         46 . The method of  claim 42 , wherein the reagent inhibits cell growth or causes apoptosis. 
     
     
         47 . The method of  claim 42 , wherein the reagent inhibits the secretion of an inflammatory cytokine. 
     
     
         48 . The method of  claim 47 , wherein the inflammatory cytokine is a tumor necrosis factor (TNF) or interleukin-1 (IL-1). 
     
     
         49 . The method of  claim 42 , wherein the reagent is an antibody that specifically binds to a polypeptide having the amino acid sequence of SEQ ID NO:2 (MKK3), SEQ ID NO:6 (MKK4-α), SEQ ID NO:8 (MKK4-β), SEQ ID NO:10 (MKK4-γ), or SEQ ID NO:4 (MKK6). 
     
     
         50 . The method of  claim 42 , wherein the reagent is a polypeptide having the amino acid sequence of SEQ ID NO:2 (MKK3), SEQ ID NO:6 (MKK4-α), SEQ ID NO:8 (MKK4-β), SEQ ID NO:10 (MKK4-γ), or SEQ ID NO:4 (MKK6), or a fragment thereof. 
     
     
         51 . The method of  claim 42 , wherein the reagent modulates MKK3 activity. 
     
     
         52 . The method of  claim 42 , wherein the reagent modulates the activity of MKK4α, MKK4β, or MKK4γ. 
     
     
         53 . The method of  claim 42 , wherein the reagent modulates MKK6 activity. 
     
     
         54 . The method of  claim 42 , wherein the reagent suppresses MKK phosphorylation of p38, JNK, or ATF2.