Effect of SB 203580 on the activity of c-Raf in vitro and in vivo

被引:129
作者
Hall-Jackson, CA
Goedert, M
Hedge, P
Cohen, P
机构
[1] Univ Dundee, Dept Biochem, MRC, Prot Phosphorylat Unit, Dundee DD1 5EH, Scotland
[2] MRC, Mol Biol Lab, Cambridge CB2 2QH, England
[3] Zeneca Pharmaceut, Macclesfield SK10 4TG, Cheshire, England
关键词
c-Raf; MAP kinase; SB203580; EGF; SAPK2; p38;
D O I
10.1038/sj.onc.1202603
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The inhibition of SAPK2a/p38 (a mitogen activated protein (MAP) kinase family member) by SE 203580 depends on the presence of threonine at residue 106. Nearly all other protein kinases are insensitive to this drug because a more bulky residue occupies this site (Eyers et al,, 1998), Raf is one of the few protein kinases that possesses threonine at this position, and we show that SE 203580 inhibits c-Raf with an IC50 of 2 mu M in vitro. However, SB 203580 does not suppress either growth factor or phorbol ester-induced activation of the classical MAP kinase cascade in mammalian cells. One of the reasons for this is that SE 203580 also triggers a remarkable activation of c-Raf in vivo (when measured in the absence of the drug). The SB203580-induced activation of c-Raf occurs without any increase in the GTP-loading of Ras, is not prevented by inhibitors of the MAPK cascade, protein kinase C or phosphatidylinositide 3-kinase, and is not triggered by the binding of this drug to SAPK2a/p38. The paradoxical activation of c-Raf by SE 203580 (and by another structurally unrelated c-Raf inhibitor) suggests that inhibitors of the kinase activity of c-Raf may not be effective as anti-cancer drugs.
引用
收藏
页码:2047 / 2054
页数:8
相关论文
共 30 条
[11]   THE INHIBITION OF GLYCOGEN-SYNTHASE KINASE-3 BY INSULIN OR INSULIN-LIKE GROWTH-FACTOR-1 IN THE RAT SKELETAL-MUSCLE CELL-LINE-L6 IS BLOCKED BY WORTMANNIN, BUT NOT BY RAPAMYCIN - EVIDENCE THAT WORTMANNIN BLOCKS ACTIVATION OF THE MITOGEN-ACTIVATED PROTEIN-KINASE PATHWAY IN L6-CELLS BETWEEN RAS AND RAF [J].
CROSS, DAE ;
ALESSI, DR ;
VANDENHEEDE, JR ;
MCDOWELL, HE ;
HUNDAL, HS ;
COHEN, P .
BIOCHEMICAL JOURNAL, 1994, 303 :21-26
[12]   SB-203580 IS A SPECIFIC INHIBITOR OF A MAP KINASE HOMOLOG WHICH IS STIMULATED BY CELLULAR STRESSES AND INTERLEUKIN-1 [J].
CUENDA, A ;
ROUSE, J ;
DOZA, YN ;
MEIER, R ;
COHEN, P ;
GALLAGHER, TF ;
YOUNG, PR ;
LEE, JC .
FEBS LETTERS, 1995, 364 (02) :229-233
[13]   Arsenite blocks growth factor induced activation of the MAP kinase cascade, upstream of Ras and downstream of Grb2-Sos [J].
Doza, YN ;
Hall-Jackson, CA ;
Cohen, P .
ONCOGENE, 1998, 17 (01) :19-24
[14]   Conversion of SB 203580-insensitive MBP kinase family members to drug-sensitive forms by a single amino-acid substitution [J].
Eyers, PA ;
Craxton, M ;
Morrice, N ;
Cohen, P ;
Goedert, M .
CHEMISTRY & BIOLOGY, 1998, 5 (06) :321-328
[15]   Activation of the Raf-1 kinase cascade by coumermycin-induced dimerization [J].
Farrar, MA ;
AlberolaIla, J ;
Perlmutter, RM .
NATURE, 1996, 383 (6596) :178-181
[16]   Activation of the novel stress-activated protein kinase SAPK4 by cytokines and cellular stresses is mediated by SKK3 (MKK6); Comparison of its substrate specificity with that of other SAP kinases [J].
Goedert, M ;
Cuenda, A ;
Craxton, M ;
Jakes, R ;
Cohen, P .
EMBO JOURNAL, 1997, 16 (12) :3563-3571
[17]   THE ACTIVATION OF DISTINCT MITOGEN-ACTIVATED PROTEIN-KINASE CASCADES IS REQUIRED FOR THE STIMULATION OF 2-DEOXYGLUCOSE UPTAKE BY INTERLEUKIN-1 AND INSULIN-LIKE GROWTH-FACTOR-I IN KB CELLS [J].
GOULD, GW ;
CUENDA, A ;
THOMSON, FJ ;
COHEN, P .
BIOCHEMICAL JOURNAL, 1995, 311 :735-738
[18]   Acquisition of sensitivity of stress-activated protein kinases to the p38 inhibitor, SB 203580, by alteration of one or more amino acids within the ATP binding pocket [J].
Gum, RJ ;
McLaughlin, MM ;
Kumar, S ;
Wang, ZL ;
Bower, MJ ;
Lee, JC ;
Adams, JL ;
Livi, GP ;
Goldsmith, EJ ;
Young, PR .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (25) :15605-15610
[19]   Effects of the inhibition of p38/RK MAP kinase on induction of five fos and jun genes by diverse stimuli [J].
Hazzalin, CA ;
Cuenda, A ;
Cano, E ;
Cohen, P ;
Mahadevan, LC .
ONCOGENE, 1997, 15 (19) :2321-2331
[20]   Amelioration of vascular dysfunctions in diabetic rats by an oral PKC beta inhibitor [J].
Ishii, H ;
Jirousek, MR ;
Koya, D ;
Takagi, C ;
Xia, P ;
Clermont, A ;
Bursell, SE ;
Kern, TS ;
Ballas, LM ;
Heath, WF ;
Stramm, LE ;
Feener, EP ;
King, GL .
SCIENCE, 1996, 272 (5262) :728-731