메뉴 건너뛰기




Volumn 20, Issue 16, 2010, Pages 4858-4864

Synthesis of potent chemical inhibitors of dynamin GTPase

Author keywords

Clathrin; Dynamin; Dynasore; Endocytosis; GTPase; Transferrin

Indexed keywords

DYNAMIN; DYNASORE; ENZYME INHIBITOR; TRANSFERRIN; UNCLASSIFIED DRUG; CLATHRIN; DD-11 COMPOUND; DD-6 COMPOUND; HYDRAZONE DERIVATIVE; N'-(3,4-DIHYDROXYBENZYLIDENE)-3-HYDROXY-2-NAPHTHAHYDRAZIDE;

EID: 77955427597     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2010.06.092     Document Type: Article
Times cited : (21)

References (26)
  • 15
    • 77955417369 scopus 로고    scopus 로고
    • note
    • 3/MeOH, 20:1) to afford the product as solid (DD-4-11).
  • 16
    • 77955417478 scopus 로고    scopus 로고
    • note
    • 3/MeOH, 20:1) to afford the product as solid (DD-12-19).
  • 17
    • 77955429846 scopus 로고    scopus 로고
    • note
    • 6. Unless otherwise noted, chemical shifts are expressed as ppm downfield from internal tetramethylsilane, or relative ppm from NMR solvent. Data are reported as follows: chemical shift, multiplicity (s, singlet; d, doublet; t, triplet; m, multiplet; b, broad; app., apparent), coupling constants, and integration. Mass spectroscopy was carried out on MALDI-TOF and LC-MS instruments.
  • 18
    • 77955421560 scopus 로고    scopus 로고
    • note
    • 3COONa, pH 4.5) and fixed in 4% paraformaldehyde. To study the reversible effect of these molecules in transferrin uptake, cells were preincubated with dynasore and derivatives (80 μM for dynasore, DD-20 and DD-13; 40 μM for DD-6, 20 μM for DD-11) for 30 min at 37 °C and washed with DPBS for various 0 min, 10 min, 30 min. After washing the cells, 20 μg/ml Texas red transferrin was treated to these cells and incubated for 10 min at 37 °C.
  • 19
    • 77955426063 scopus 로고    scopus 로고
    • Note
    • Images were obtained with an Olympus IX-71 inverted microscope (Olympus Optical, Tokyo, Japan) with a 40×, 1.0 N.A. oil lens using a CoolSNAP-Hq CCD camera (Roper Scientific, Tucson, AZ) driven by MetaMorph imaging software (Universal Imaging Corporation, West Chester, PA) with a Texas red optimized filter set (Omega Optical, Brattleboro, VT). Light from a mercury lamp was shuttered using a VMM1 Uniblitz shutter (Vincent Associates, Rochester, NY). Analysis and quantification of data were performed using MetaMorph software and SigmaPlot 8.0 (Systat Software, Point Richmond, CA), and data were presented as mean ± SE.
  • 20
    • 77955425087 scopus 로고    scopus 로고
    • note
    • 50 by nonlinear regression curve fitting using Graphad Prism 5 software. GTPase activity of 1 μM Drp1 was measured by same method with dynamin I GTPase activity. To identify the effect of dynasore derivatives to Cdc42 GTPase activity, 1 μM Cdc42 (Cytoskeleton) was mixed with 0.5 mM GST tagged human SNX26 RhoGAP domain, and added to prepared ELIPA mixture with 1 mM GTP. The amount of inorganic phosphate released by GTPase activity was measured by using the GTPase Enzyme Linked Inorganic Phosphate Assay in the presence of 0.5 μM GST tagged human TCGAP RhoGAP domain. The amount of inorganic phosphate released by Cdc42 in the presence of 0.1% DMSO (control), 80 μM dynaosre, DD-6 or DD-11 was measured at 360 nm and plotted as a function of time.
  • 21
    • 77955413810 scopus 로고    scopus 로고
    • note
    • M software (Olympus). Laser lines (488 and 561 nm diode lasers) were coupled to the TIRFM condenser through two independent optical fibers. The calculated evanescent depth was ≈150 nm. Cells were typically imaged in two channels by sequential excitation with 0.15 s exposures and detected with back-illuminated Andor iXon887 EMCCD camera (512 × 512, 16-bit; Andor Technologies). Image J program (National Institutes of Health) was used for analysis.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.