Identification of MAPKAP kinase 2 as a major enzyme responsible for the phosphorylation of the small mammalian heat shock proteins

MAP kinase‐activated protein kinase‐2 (MAPKAP kinase‐2) phosphorylates the serine residues in murine heat shock protein 25 (hsp25) and human heat shock protein 27 (hsp27) which are phosphorylated in vivo in response to growth factors and heat shock, namely Ser15 and Ser40 (hsp25) and Ser15, Ser78 and Ser82 (hsp27). Se86 of hsp25 and the equivalent residue in hsp27 (Ser82) are phosphorylated preferentially in vitro. The small heat shock protein is present in rabbit skeletal muscle and hsp25 kinase activity in skeletal muscle extracts co‐purifies with MAPKAP kinase‐2 activity throughout the purification of the latter enzyme. These results suggest that MAPKAP kinase‐2 is the enzyme responsible for the phosphorylation of these small heat shock proteins in mammalian cells.

[1]  F. Guesdon,et al.  Phosphorylation of the small heat-shock protein is regulated by interleukin 1, tumour necrosis factor, growth factors, bradykinin and ATP. , 1991, The Biochemical journal.

[2]  R. Kraft,et al.  Molecular cloning, sequencing and expression in Escherichia coli of the 25-kDa growth-related protein of Ehrlich ascites tumor and its homology to mammalian stress proteins. , 1989, European journal of biochemistry.

[3]  R. Fletterick,et al.  Human muscle glycogen synthase cDNA sequence: a negatively charged protein with an asymmetric charge distribution. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[4]  J. Saklatvala,et al.  Interleukin 1 and tumour necrosis factor increase phosphorylation of fibroblast proteins , 1988, FEBS letters.

[5]  T. Sturgill,et al.  Recent progress in characterization of protein kinase cascades for phosphorylation of ribosomal protein S6. , 1991, Biochimica et biophysica acta.

[6]  C. Anderson,et al.  Human HSP27 is phosphorylated at serines 78 and 82 by heat shock and mitogen-activated kinases that recognize the same amino acid motif as S6 kinase II. , 1992, The Journal of biological chemistry.

[7]  W. Fiers,et al.  Rapid phosphorylation of a 27 kDa protein induced by tumor necrosis factor , 1988, FEBS letters.

[8]  M. Gaestel,et al.  Over‐expression of the small heat‐shock protein, hsp25, inhibits growth of Ehrlich ascites tumor cells , 1992, FEBS letters.

[9]  H. Tung,et al.  Protein phosphatase-1 and protein phosphatase-2A from rabbit skeletal muscle. , 1988, Methods in enzymology.

[10]  W. Welch,et al.  Characterization and purification of the small 28,000-dalton mammalian heat shock protein. , 1987, The Journal of biological chemistry.

[11]  P. Dent,et al.  The molecular mechanism by which insulin stimulates glycogen synthesis in mammalian skeletal muscle , 1990, Nature.

[12]  W. Welch Phorbol ester, calcium ionophore, or serum added to quiescent rat embryo fibroblast cells all result in the elevated phosphorylation of two 28,000-dalton mammalian stress proteins. , 1985, The Journal of biological chemistry.

[13]  G. Crabtree,et al.  Rapamycin-FKBP specifically blocks growth-dependent activation of and signaling by the 70 kd S6 protein kinases , 1992, Cell.

[14]  P. Cohen,et al.  MAPKAP kinase‐2; a novel protein kinase activated by mitogen‐activated protein kinase. , 1992, The EMBO journal.

[15]  V. Erdmann,et al.  Identification of the phosphorylation sites of the murine small heat shock protein hsp25. , 1991, The Journal of biological chemistry.

[16]  P. Cohen,et al.  Purification and characterisation of the insulin-stimulated protein kinase from rabbit skeletal muscle; close similarity to S6 kinase II. , 1991, European journal of biochemistry.

[17]  J. Buchner,et al.  Hsp90 chaperones protein folding in vitro , 1992, Nature.

[18]  D. Fabbro,et al.  The 27,000 daltons stress proteins are phosphorylated by protein kinase C during the tumor promoter-mediated growth inhibition of human mammary carcinoma cells. , 1988, Biochemical and biophysical research communications.

[19]  P. Cohen,et al.  Substrate specificity of a multifunctional calmodulin-dependent protein kinase. , 1985, The Journal of biological chemistry.

[20]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[21]  J. Landry,et al.  Heat shock resistance conferred by expression of the human HSP27 gene in rodent cells , 1989, The Journal of cell biology.

[22]  E. Krebs,et al.  Effect of proteases on the structure and activity of rabbit skeletal muscle glycogen synthetase , 1979, FEBS letters.

[23]  J. Maller,et al.  Purification and characterization of a protein kinase from Xenopus eggs highly specific for ribosomal protein S6. , 1986, The Journal of biological chemistry.

[24]  P. Chrétien,et al.  Enhanced constitutive expression of the 27‐kDa heat shock proteins in heat‐resistant variants from chinese hamster cells , 1988, Journal of cellular physiology.

[25]  J. Maller,et al.  Insulin-stimulated MAP-2 kinase phosphorylates and activates ribosomal protein S6 kinase II , 1988, Nature.