Journal: Cell Communication and Signaling : CCS

Article Title: Identification of a selective inhibitor of IDH2/R140Q enzyme that induces cellular differentiation in leukemia cells

doi: 10.1186/s12964-020-00536-7

Figure Lengend Snippet: In vitro inhibition of the inhibitors with homodimeric IDH enzymes

Article Snippet: The human recombinant C-terminal FLAG-tag IDH enzymes including IDH2/R140Q, IDH2/WT, IDH1/R132H and IDH1/WT homodimer were purchased from BPS Bioscience (San Diego, USA).

Techniques: In Vitro, Inhibition, Incubation

Journal: Cell Communication and Signaling : CCS

Article Title: Identification of a selective inhibitor of IDH2/R140Q enzyme that induces cellular differentiation in leukemia cells

doi: 10.1186/s12964-020-00536-7

Figure Lengend Snippet: Identification of CP-17 as an inhibitor of IDH2/R140Q. a Structure-based in silico screening of a small molecule library (ChemDiv) for compounds that bind to the allosteric site of IDH2/R140Q identifies a hit compound CP-17 with IC 50 of 40.75 nM. HTVS: High-throughput Virtual Screening mode; SP: Standard-Precision mode; XP: Extra-Precision mode. b Structure of CP-17 binding at the allosteric site of IDH2/R140Q. The protein was represented as cartoon (colored in light blue and yellow for each monomer), the carbon atoms of CP-17 as magenta sticks, NADPH and Q140 as green sticks, and the Ca 2+ as black spheres. c Molecular interactions of CP-17 binding at the IDH2/R140Q dimer interface (light blue and yellow for carbon atoms of adjacent monomer residues). The hydrogen bonds between CP-17 and Q316 residue were labeled with red dotted lines

Article Snippet: The human recombinant C-terminal FLAG-tag IDH enzymes including IDH2/R140Q, IDH2/WT, IDH1/R132H and IDH1/WT homodimer were purchased from BPS Bioscience (San Diego, USA).

Techniques: In Silico, High Throughput Screening Assay, Binding Assay, Labeling

Journal: Cell Communication and Signaling : CCS

Article Title: Identification of a selective inhibitor of IDH2/R140Q enzyme that induces cellular differentiation in leukemia cells

doi: 10.1186/s12964-020-00536-7

Figure Lengend Snippet: The calculated binding free energy of NADPHs with IDH2/R140Q in different complexes

Article Snippet: The human recombinant C-terminal FLAG-tag IDH enzymes including IDH2/R140Q, IDH2/WT, IDH1/R132H and IDH1/WT homodimer were purchased from BPS Bioscience (San Diego, USA).

Techniques: Binding Assay

Journal: Cell Communication and Signaling : CCS

Article Title: Identification of a selective inhibitor of IDH2/R140Q enzyme that induces cellular differentiation in leukemia cells

doi: 10.1186/s12964-020-00536-7

Figure Lengend Snippet: Molecular dynamics (MD) simulations of IDH2/R140Q_CP-17, IDH2/R140Q_α-KG and IDH2/WT_CP-17 systems. a Evolution of RMSDs during 200 ns MD simulations. b Evolution of the Ile116-Leu289′ distances and the Ile116-Phe148-Leu289′ angles (measured on Cα atoms) in monomers A and B during MD simulations

Article Snippet: The human recombinant C-terminal FLAG-tag IDH enzymes including IDH2/R140Q, IDH2/WT, IDH1/R132H and IDH1/WT homodimer were purchased from BPS Bioscience (San Diego, USA).

Techniques:

Journal: Cell Communication and Signaling : CCS

Article Title: Identification of a selective inhibitor of IDH2/R140Q enzyme that induces cellular differentiation in leukemia cells

doi: 10.1186/s12964-020-00536-7

Figure Lengend Snippet: The average distances of Ile116-Leu289’ and angles of Ile116-Phe148-Leu289’ between Cα atoms calculated from the 100–200 ns MD simulation trajectory

Article Snippet: The human recombinant C-terminal FLAG-tag IDH enzymes including IDH2/R140Q, IDH2/WT, IDH1/R132H and IDH1/WT homodimer were purchased from BPS Bioscience (San Diego, USA).

Techniques:

Journal: Cell Communication and Signaling : CCS

Article Title: Identification of a selective inhibitor of IDH2/R140Q enzyme that induces cellular differentiation in leukemia cells

doi: 10.1186/s12964-020-00536-7

Figure Lengend Snippet: Conformational changes of IDH2/WT bound with CP-17 after MD simulation. RMSD matrices of IDH2/R140Q_CP-17 a and IDH2/WT_CP-17 b systems during MD simulation. c Representation of surface maps of CP-17 bound IDH2/WT structure, with the catalytic sites of monomer A in an active closed conformation (left) and monomer B in an inactive open conformation (right)

Article Snippet: The human recombinant C-terminal FLAG-tag IDH enzymes including IDH2/R140Q, IDH2/WT, IDH1/R132H and IDH1/WT homodimer were purchased from BPS Bioscience (San Diego, USA).

Techniques:

Journal: Cell Communication and Signaling : CCS

Article Title: Identification of a selective inhibitor of IDH2/R140Q enzyme that induces cellular differentiation in leukemia cells

doi: 10.1186/s12964-020-00536-7

Figure Lengend Snippet: CP-17 treatment reversed GM-CSF-independent growth induced by IDH2/R140Q. IDH2/R140Q expression confers GM-CSF independent growth of the TF-1 cells. Proliferation of TF-1(IDH2/R140Q) ( a ) and TF-1(WT) ( b ) cells with the treatment of CP-17. (* P < 0.01 vs 7 day of Control, # P < 0.01 vs 0 day of Control). The variance is similar between the groups that are being statistically compared

Article Snippet: The human recombinant C-terminal FLAG-tag IDH enzymes including IDH2/R140Q, IDH2/WT, IDH1/R132H and IDH1/WT homodimer were purchased from BPS Bioscience (San Diego, USA).

Techniques: Expressing

Journal: Cell Communication and Signaling : CCS

Article Title: Identification of a selective inhibitor of IDH2/R140Q enzyme that induces cellular differentiation in leukemia cells

doi: 10.1186/s12964-020-00536-7

Figure Lengend Snippet: CP-17 reversed the TF-1(IDH2/R140Q) differentiation block. a IC 50 value of intracellular D-2-HG inhibition by CP-17 treatment in TF-1(IDH2/R140Q) cells. b TF-1(WT) or TF-1(IDH2/R140Q) cells were induced with 50 ng/mL EPO to differentiate for 7 days in the presence of 0, 1 μM, and 3 μM of CP-17. After then cells were collected, and color change induced by hemoglobin γ expression was photographed. D-2-HG level and hemoglobin γ protein expression were detected with LC-MS and western blot respectively

Article Snippet: The human recombinant C-terminal FLAG-tag IDH enzymes including IDH2/R140Q, IDH2/WT, IDH1/R132H and IDH1/WT homodimer were purchased from BPS Bioscience (San Diego, USA).

Techniques: Blocking Assay, Inhibition, Expressing, Liquid Chromatography with Mass Spectroscopy, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: Identification of a selective inhibitor of IDH2/R140Q enzyme that induces cellular differentiation in leukemia cells

doi: 10.1186/s12964-020-00536-7

Figure Lengend Snippet: CP-17 decreased high level of histone methylation in TF-1(IDH2/R140Q) cells. TF-1(IDH2/R140Q) and TF-1(WT) cells were treated with CP-17 (1 μM, 3 μM) for 7 days, after which expression levels of H3K4Me3, H3K9Me3, H3K27Me3 and H3 were assessed by western blot with specific antibodies. The total amount of H3 was used as loading control

Article Snippet: The human recombinant C-terminal FLAG-tag IDH enzymes including IDH2/R140Q, IDH2/WT, IDH1/R132H and IDH1/WT homodimer were purchased from BPS Bioscience (San Diego, USA).

Techniques: Methylation, Expressing, Western Blot