Identification of a novel small molecule targeting UQCRB of mitochondrial complex III and its anti-angiogenic activity

Bioorganic and Medicinal Chemistry Letters

Volumn 21, Issue 3, 2011, Pages 1052-1056

  Jung, Hye Jin ^a   Kim, Ki Hyun ^a   Kim, Nam Doo ^b   Han, Gyoonhee ^a   Kwon, Ho Jeong ^a  

^a Yonsei University   (South Korea)

^b Equispharm Co Ltd   (South Korea)

Author keywords

Angiogenesis; HDNT; O2 sensing; UQCRB

Indexed keywords

6 [(1 HYDROXYNAPHTHALEN 4 YLAMINO)DIOXYSULFONE] 2H NAPHTHO[1,8 BC]THIOPHEN 2 ONE; ANGIOGENESIS INHIBITOR; REACTIVE OXYGEN METABOLITE; UBIQUINOL CYTOCHROME C REDUCTASE; UNCLASSIFIED DRUG;

ANTIANGIOGENIC ACTIVITY; ARTICLE; CONTROLLED STUDY; CYTOTOXICITY; DRUG IDENTIFICATION; DRUG PROTEIN BINDING; DRUG STRUCTURE; GENE TARGETING; HUMAN; HUMAN CELL; HYPOXIA; IC 50; IN VITRO STUDY; MOLECULAR WEIGHT; OXYGEN SENSING; PROTEIN FUNCTION; SIGNAL TRANSDUCTION; UMBILICAL VEIN ENDOTHELIAL CELL;

ANGIOGENESIS INHIBITORS; BINDING SITES; CARRIER PROTEINS; CELL HYPOXIA; CELLS, CULTURED; COMPUTER SIMULATION; ELECTRON TRANSPORT COMPLEX III; ENDOTHELIAL CELLS; HETEROCYCLIC COMPOUNDS, 3-RING; HUMANS; MITOCHONDRIA; REACTIVE OXYGEN SPECIES; SIGNAL TRANSDUCTION; SULFONAMIDES;

EID: 78751642080     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2010.12.002     Document Type: Article

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20. 7 pfu/mL) and added to the terpestacin-immobilized wells in the presence or absence of competitors (100 μM). After incubation for 1 h at room temperature with gentle shaking, the well plate was washed with TBS and bound phage particles were eluted with 100 μM terpestacin diluted in TBS. The binding phages were infected into the Escherichia coli strain BLT5615 grown on Luria-Bertani (LB) agar plates supplemented with 50 μg/mL ampicillin, and then the plaques were counted to calculate plaque forming units (pfus).
21. Competitive binding assay by fluorescence staining: HUVECs were incubated with 20 μM coumarin or coumarin-conjugated terpestacin (ter-coumarin) for 24 h in the presence or absence of competitors (10 μM HDNT or 30 μM terpestacin). After washing with phosphate-buffered saline (PBS, pH 7.4), cells were observed under a fluorescence microscope (Olympus). The fluorescence intensity was measured using an FL600 microplate fluorescence reader (Bio-Tek).
22. Competitive binding assay by SPR: Biotin and biotinylated terpestacin were sequentially immobilized onto the surface of a streptavidin-coated sensor chip. Competitors (25 μM) were incubated with purified UQCRB protein (25 μM) in the running buffer (10 mM HEPES, 150 mM NaCl, 3 mM EDTA (pH 7.4)), and then UQCRB-competitor complexes were injected onto the sensor chip at a flow rate of 30 μL/min. The surface of the sensor chip was regenerated by injection of 5 μL of the regeneration buffer (50 mM NaOH). Molecular interaction analysis was performed using the BIAcore 2000 system (BIAcore AB) and the apparent binding affinities were calculated by subtraction of resonance values of UQCRB binding to control biotin from those of UQCRB binding to biotinylated terpestacin.
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