Tyrosine phosphorylation is crucial for growth signaling by tissue inhibitors of metalloproteinases (TIMP-1 and TIMP-2).
[3H]Thymidine (TdR) incorporation by human osteosarcoma cell line MG-63 was significantly stimulated at as early as 3 h after the addition of either TIMP-1 or TIMP-2 alone. Maximum stimulation was attained at a concentration of either 20 ng/ml (0.71 nM) TIMP-1 or 1.0 ng/ml (46 pM) TIMP-2. Tyrosine kinase inhibitors such as genistein, erbstatin, and herbimycin A almost completely inhibited the [3H]TdR incorporation stimulated by either of the TIMPs. However, essentially no effect was observed with H-89, H-7, bisindolylmaleimide and K-252a. These inhibition studies suggest a crucial role for tyrosine kinase in the signal transduction of TIMPs. Phosphotyrosine-containing proteins were significantly elevated by the treatment with both TIMPs. We also found that either TIMP stimulated an increase in mitogen-activated protein (MAP) kinase activity, suggesting that MAP kinase plays a role in TIMP-dependent growth signaling.
Department of Biochemistry, School of Dentistry, Aichi-Gakuin University, Nagoya, Japan.
K
K
Yamashita
M
M
Suzuki
H
H
Iwata
T
T
Koike
M
M
Hamaguchi
A
A
Shinagawa
T
T
Noguchi
T
T
Hayakawa
D016227:Benzoquinones; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D004247:DNA / Q000096:biosynthesis / Q000187:drug effects; D004789:Enzyme Activation; D019833:Genistein; D006023:Glycoproteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006801:Humans; D006873:Hydroquinones / Q000378:metabolism / Q000494:pharmacology; D007529:Isoflavones / Q000378:metabolism / Q000494:pharmacology; D047029:Lactams, Macrocyclic; D019950:Mitogen-Activated Protein Kinase 1; D012516:Osteosarcoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D010766:Phosphorylation; D011480:Protease Inhibitors / Q000378:metabolism / Q000494:pharmacology*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011506:Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011809:Quinones / Q000378:metabolism / Q000494:pharmacology; D017828:Rifabutin / Q000031:analogs & derivatives; D015398:Signal Transduction; D013936:Thymidine / Q000378:metabolism / Q000493:pharmacokinetics; D013997:Time Factors; D019716:Tissue Inhibitor of Metalloproteinase-2; D019714:Tissue Inhibitor of Metalloproteinases; D014316:Tritium; D014407:Tumor Cells, Cultured; D014443:Tyrosine / Q000378:metabolism*
D016227:Benzoquinones; D006023:Glycoproteins; D006873:Hydroquinones; D007529:Isoflavones; D047029:Lactams, Macrocyclic; D011480:Protease Inhibitors; D011506:Proteins; D011809:Quinones; D019714:Tissue Inhibitor of Metalloproteinases; D014316:Tritium; D019716:Tissue Inhibitor of Metalloproteinase-2; D017828:Rifabutin; D014443:Tyrosine; C020754:herbimycin; D004247:DNA; D019833:Genistein; D011505:Protein-Tyrosine Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D019950:Mitogen-Activated Protein Kinase 1; D013936:Thymidine; C048212:erbstatin