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Volumn 19, Issue 8, 2009, Pages 2230-2234

1-Aryl-3,4-dihydroisoquinoline inhibitors of JNK3

Author keywords

Inhibitor; JNK; JNK3; Kinase; MAPK; Selective

Indexed keywords

3,4 DIHYDROISOQUINOLINE DERIVATIVE; ISOQUINOLINE DERIVATIVE; MITOGEN ACTIVATED PROTEIN KINASE 1; MITOGEN ACTIVATED PROTEIN KINASE P38; STRESS ACTIVATED PROTEIN KINASE INHIBITOR; UNCLASSIFIED DRUG;

EID: 63149148930     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2009.02.098     Document Type: Article
Times cited : (35)

References (39)
  • 14
    • 63149183873 scopus 로고    scopus 로고
    • note
    • 50 > 6.
  • 15
    • 63149122988 scopus 로고    scopus 로고
    • note
    • 2 (5 mM final) in assay buffer (40 mM HEPES pH 7.4, 1 mM DTT) were added to wells containing 1 μl of various concentrations of compound or DMSO vehicle (3% final) in NUNC 384-well black plates. The reaction was initiated by addition of p38α (100 pM final) to give a total volume of 30 μl. After 120 min incubation (rt), 15 μl of 100 mM EDTA pH 7.4 was added followed by detection reagent (15 μl) in buffer (100 mM HEPES pH 7.4, 150 mM NaCl, 0.1% w/v/BSA, 1 mM DTT) containing antiphosphothreonine-ATF2-71 polyclonal antibody (Cell Signalling Technology, Beverly Massachusetts, MA) labelled with W-1024 Eu chelate (Wallac OY, Turku, Finland), and APC-labelled streptavidin (Prozyme, San Leandro, CA). After 60 min further incubation (rt) the ATF-2 phosphorylation was measured using a Packard Discovery plate reader (Perkin-Elmer, Pangbourne, UK) as a ratio of specific 665 nm energy transfer signal to reference Eu 620 nm signal.
  • 16
    • 63149113437 scopus 로고    scopus 로고
    • note
    • 18
  • 20
    • 63149154322 scopus 로고    scopus 로고
    • note
    • TM service, for further details see: www.millipore.com. JNK data were obtained using N-terminal His-tagged full-length human JNK1α1, JNK2α2 or JNK3 with ATF2 as substrate, in the presence of 45, 45, or 10 μM ATP, respectively. Erk-2 data were obtained in a radiometric filter binding assay using N-terminal GST-tagged full-length human Erk-2, activated with MEK1, with myelin basic protein as substrate, in the presence of 155 μM ATP.
  • 21
    • 63149165984 scopus 로고    scopus 로고
    • note
    • 14 was purified following a five stage process after lysis of E. coli cells expressing GST-JNK3t. [Glutathione Sepharose (GSH), Thrombin cleavage, GSH, Source 15-Q anion exchange, SEC]. The protein was supplied in 50 mM Tris/HCl pH 8.0, 150 mM NaCl post final stage Superdex 200 prep grade Size Exclusion column. Co-crystals of Jnk3t with 16 were grown at 20 °C using the hanging drop method combined with micro-seeding. Protein at 13 mg/mL, pre-incubated with 5 mM compound, was mixed with serial dilutions of a Jnk3t seed stock (made in 25% peg 3350, 0.1 M sodium Hepes pH7.5). Drops were then equilibrated over a reservoir containing 18% peg3350, 0.1 M sodium Hepes pH7.5) before freezing in mother liquor plus 15% glycerol. A 2.4 Å dataset was collected from a single frozen crystal of Jnk3t/16 on a Mar345 detector mounted on a micromax 007HF rotating anode generator. The crystal structure was refined starting from the coordinates of another Jnk3t complex crystal stucture (pdb code: 2O0U-ligand removed before start of refinement). The deposition code for the Jnk3t/16 complex is 2waj. Crystal structures of 24 and 26 with a similarly truncated version of Jnk1 are virtually identical (Bax et al., unpublished results).
  • 26
    • 63149086135 scopus 로고    scopus 로고
    • note
    • Note that the hydrogen atoms were not resolved in this structure: where necessary for analysis of the H-bond geometries they were added in standard positions using Maestro (Schrödinger Inc.).
  • 27
    • 0003914038 scopus 로고    scopus 로고
    • Jeffrey G.A. (Ed), Oxford University Press, New York
    • In: Jeffrey G.A. (Ed). An Introduction to Hydrogen Bonding (1997), Oxford University Press, New York
    • (1997) An Introduction to Hydrogen Bonding


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.