Novel compounds, '1,3-selenazine derivatives' as specific inhibitors of eukaryotic elongation factor-2 kinase

被引:65
作者
Cho, SI
Koketsu, I
Ishihara, H
Matsushita, M
Nairn, AC
Fukazawa, H
Uehara, Y
机构
[1] Natl Inst Infect Dis, Dept Bioact Mol, Shinjuku Ku, Tokyo 1628640, Japan
[2] Gifu Univ, Fac Engn, Dept Chem, Gifu 5011193, Japan
[3] Okayama Univ, Sch Med, Dept Physiol 1, Okayama 7008558, Japan
[4] Rockefeller Univ, Mol & Cellular Neurosci Lab, New York, NY 10021 USA
来源
BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS | 2000年 / 1475卷 / 03期
关键词
1,3-selenazine derivative; rottlerin; eukaryotic elongation factor-2 kinase; specific eukaryotic elongation factor-2 kinase inhibitor; calmodulin-dependent kinase III;
D O I
10.1016/S0304-4165(00)00061-1
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 inhibitory activities of 5,6-dihydro-4H-1,3-selenazine derivatives on protein kinases were investigated. In a multiple protein kinase assay using a postnuclear fraction of v-src-transformed NIH3T3 cells, 4-ethyl-3-hydroxy-2-p-tolyl-5,6-dihydro-4H-1,3-selenazine (TS-2) and 4-hydroxy-6-isopropyl-4-methyl-2-p-tolyl-5,6-dihydro-4H-1,3-selenazine (TS-4) exhibited selective inhibitory activity against eukaryotic elongation factor-2 kinase (eEF-2K) over protein kinase A (PKA), protein kinase C (PKC) and protein tyrosine kinase (PTK). In further experiments using purified kinases. TS-2 (IC50 = 0.36 mu M) and TS-4 (IC50 = 0.31 mu M) inhibited eEF-2K about 25-fold more effectively than calmodulin-dependent protein kinase-I (CaMK-I), and about 6-fold (TS-2) or 33-fold (TS-4) more effectively than calmodulin-dependent protein kinase-II (CaMK-II), respectively. TS-2 and TS-4 showed much weaker inhibitory activity toward PKA and PKC, while TS-4, but not TS-2, moderately inhibited immunoprecipitated v-src kinase. TS-2 (10.7-fold) and TS-4 (12.5-fold) demonstrated more potent and more specific eEF-2K inhibitory activity than rottlerin, a previously identified eEF-2K inhibitor. TS-2 inhibited ATP or eEF-2 binding to eEF-2K in a competitive or non-competitive manner, respectively. In cultured v-si c-transformed NIH3T3 cells, TS-2 also decreased phospho-eEF-2 protein level (IC50 = 4.7 mu M) without changing the total eEF-2 protein level. Taken together, these results suggest that TS-2 and TS-4 are the first identified selective eEF-2K inhibitors and should be useful tools for studying the function of eEF-2K. (C) 2000 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:207 / 215
页数:9
相关论文
共 56 条
[1]   Kinase cascades regulating entry into apoptosis [J].
Anderson, P .
MICROBIOLOGY AND MOLECULAR BIOLOGY REVIEWS, 1997, 61 (01) :33-+
[2]  
BAGAGLIO DM, 1993, CANCER RES, V53, P2260
[3]   Leukocyte protein tyrosine kinases: Potential targets for drug discovery [J].
Bolen, JB ;
Brugge, JS .
ANNUAL REVIEW OF IMMUNOLOGY, 1997, 15 :371-404
[4]   NERVE GROWTH FACTOR-INDUCED DOWN-REGULATION OF CALMODULIN-DEPENDENT PROTEIN KINASE-III IN PC12 CELLS INVOLVES CYCLIC AMP-DEPENDENT PROTEIN-KINASE [J].
BRADY, MJ ;
NAIRN, AC ;
WAGNER, JA ;
PALFREY, HC .
JOURNAL OF NEUROCHEMISTRY, 1990, 54 (03) :1034-1039
[5]  
CARPENTER G, 1978, NATURE, V276, P409, DOI 10.1038/276409a0
[6]   INCREASED PHOSPHORYLATION OF ELONGATION FACTOR-II DURING MITOSIS IN TRANSFORMED HUMAN AMNION CELLS CORRELATES WITH A DECREASED RATE OF PROTEIN-SYNTHESIS [J].
CELIS, JE ;
MADSEN, P ;
RYAZANOV, AG .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (11) :4231-4235
[7]  
COHEN VI, 1979, SYNTHESIS-STUTTGART, P66
[8]   Mapping of the novel protein kinase catalytic domain of Dictyostelium myosin II heavy chain kinase A [J].
Cote, GP ;
Luo, X ;
Murphy, MB ;
Egelhoff, TT .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (11) :6846-6849
[9]   Analysis of the domain structure of elongation factor-2 kinase by mutagenesis [J].
Diggle, TA ;
Seehra, CK ;
Hase, S ;
Redpath, NT .
FEBS LETTERS, 1999, 457 (02) :189-192
[10]   REGULATORY MECHANISMS IN THE CONTROL OF PROTEIN-KINASES [J].
FLOCKHART, DA ;
CORBIN, JD .
CRC CRITICAL REVIEWS IN BIOCHEMISTRY, 1982, 12 (02) :133-186