Targeting Ovarian Tumor Cell Adhesion Mediated by Tissue Transglutaminase

被引:31
作者
Khanna, May [1 ]
Chelladurai, Bhadrani [2 ]
Gavini, Aruna [2 ]
Li, Liwei [1 ,6 ]
Shao, Minghai [2 ]
Courtney, David [2 ]
Turchi, John J. [1 ,2 ,5 ]
Matei, Daniela [1 ,3 ,5 ]
Meroueh, Samy [1 ,4 ,5 ,6 ,7 ]
机构
[1] Indiana Univ Sch Med, Dept Biochem & Mol Biol, Indianapolis, IN 46202 USA
[2] Indiana Univ Sch Med, Dept Med, Indianapolis, IN 46202 USA
[3] Indiana Univ Sch Med, Dept Obstet & Gynecol, Indianapolis, IN 46202 USA
[4] Indiana Univ Sch Med, Dept Chem & Chem Biol, Indianapolis, IN 46202 USA
[5] Indiana Univ Sch Med, IU Simon Canc Ctr, Indianapolis, IN 46202 USA
[6] Indiana Univ Sch Med, Ctr Computat Biol & Bioinformat, Indianapolis, IN 46202 USA
[7] Indiana Univ Sch Med, Stark Neurosci Res Inst, Indianapolis, IN 46202 USA
关键词
PROMOTE PROTEIN STABILITY; CANCER METASTASIS; BINDING; FIBRONECTIN; IDENTIFICATION; SPHEROIDS; MECHANISM; MIGRATION; COLLAGEN; DOMAIN;
D O I
10.1158/1535-7163.MCT-10-0912
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Tissue transglutaminase (TG2) is a transpeptidase involved in protein cross-linking through generation of epsilon-(gamma-glutamyl) lysine isopeptide bonds. It also promotes cell adhesion through interaction with fibronectin and facilitates formation of fibronectin-integrin complexes. This interaction is involved in tumor cell adhesion to the matrix and in the process of tumor dissemination. Its inhibition by small molecules may therefore be useful in blocking metastasis. To that end, we screened more than 800,000 compounds following an in silico docking approach targeting two distinct cavities in the vicinity of the fibronectin-binding site on TG2. A total of 120 compounds were acquired and tested in cell culture-based assays for inhibition of ovarian tumor cell adhesion and proliferation. Seven compounds showed more than 50% inhibition of cell adhesion at a concentration of 25 mu mol/L. A follow-up fluorescence polarization study revealed that one compound in particular (ITP-79) inhibited binding of a TG2 peptide to a 42-kDa fragment of fibronectin in a dose-dependent manner. This inhibition was confirmed in cancer cells by coimmunoprecipitation. A competition assay with surface plasmon resonance showed that ITP-79 modulated binding of TG2 to fibronectin. Direct binding of compounds that inhibited adhesion to TG2 were examined with differential scanning fluorimetry, which measures the effect of the compound on the melting temperature of the target. Two compounds, including ITP-79, reduced TG2 stabilization, mimicking the effects of GTP, a known negative allosteric regulator of TG2 enzymatic function. This suggests a potential allosteric mechanism for the compound in light of its distal target site. Mol Cancer Ther; 10(4); 626-36. (C)2011 AACR.
引用
收藏
页码:626 / 636
页数:11
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