Identification of small molecules that inhibit the interaction of TEM8 with anthrax protective antigen using a FRET assay.
Identification of small molecules that inhibit the interaction of TEM8 with anthrax protective antigen using a FRET assay.
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DOI：
10.1177/1087057113478655
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发表时间：
2013-07
影响因子：
--
通讯作者：
Rogers MS
中科院分区：
文献类型：
--
作者：
Cryan LM;Habeshian KA;Caldwell TP;Morris MT;Ackroyd PC;Christensen KA;Rogers MS
肿瘤标志物内皮8(TEM8)是炭疽毒素保护性抗原(PA)的受体。与正常血管相比，TEM8在肿瘤血管内皮细胞上表达上调。许多研究表明，TEM8在发育和肿瘤血管生成中起着关键作用。我们还表明，以突变形式的PA靶向炭疽受体可以抑制体内血管生成和肿瘤形成。在这里，我们描述了高通量荧光共振能量转移分析的发展和测试，以确定强烈抑制PA和TEM8相互作用的分子。我们描述的分析是灵敏和稳健的，Z素值为0.8。初步筛选了2310个已知的生物活性文库化合物，确定ebselen和硫柳汞是TEM8-PA相互作用的抑制剂。这些分子都含有半胱氨酸-反应性过渡金属，补充研究表明，它们相互作用的抑制是由于TEM8胞外区的半胱氨酸残基的修饰。这是鉴定TEM8抑制剂的高通量筛选试验的第一次展示，具有抗炭疽和抗血管生成疾病的潜在应用。
Tumor marker endothelial 8 (TEM8) is a receptor for the Protective Antigen (PA) component of anthrax toxin. TEM8 is upregulated on endothelial cells lining the blood vessels within tumors, compared to normal blood vessels. A number of studies have demonstrated a pivotal role for TEM8 in developmental and tumor angiogenesis. We have also shown that targeting the anthrax receptors with a mutated form of PA inhibits angiogenesis and tumor formation in vivo. Here we describe the development and testing of a high-throughput fluorescence resonance energy transfer assay to identify molecules that strongly inhibit the interaction of PA and TEM8. The assay we describe is sensitive and robust, with a Z-prime value of 0.8. A preliminary screen of 2310 known bioactive library compounds identified ebselen and thimerosal as inhibitors of the TEM8-PA interaction. These molecules each contain a cysteine-reactive transition metal, and complimentary studies indicate that their inhibition of interaction is due to modification of a cysteine residue in the TEM8 extracellular domain. This is the first demonstration of a high-throughput screening assay that identifies inhibitors of TEM8, with potential application for anti-anthrax and anti-angiogenic diseases.
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DOI：
10.1073/pnas.0431098100
发表时间：
2003-04-29
影响因子：
11.1
作者：
Scobie, HM;Rainey, GJA;Young, JAT
通讯作者：
Young, JAT
影响因子：
82.9
作者：
通讯作者：
--
影响因子：
11.2
作者：
Rogers, Michael S.;Christensen, Kenneth A.;D'Amato, Robert J.
通讯作者：
D'Amato, Robert J.
影响因子：
11.2
作者：
Cullen M;Seaman S;Chaudhary A;Yang MY;Hilton MB;Logsdon D;Haines DC;Tessarollo L;St Croix B
通讯作者：
St Croix B
影响因子：
6.4
作者：
Scobie, HM;Thomas, D;Manchester, M
通讯作者：
Manchester, M