A rapid and sensitive high-throughput screening method to identify compounds targeting protein-nucleic acids interactions.
A rapid and sensitive high-throughput screening method to identify compounds targeting protein-nucleic acids interactions.
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DOI：
10.1093/nar/gkv069
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发表时间：
2015-04-30
影响因子：
14.9
通讯作者：
Leng F
中科院分区：
文献类型：
--
作者：
Alonso N;Guillen R;Chambers JW;Leng F
dna结合蛋白和rna结合蛋白通常被认为是“不可药物的”，部分原因是缺乏一种有效的方法来从现有的小分子库中识别抑制剂。本文报道了一种基于蛋白质- dna或蛋白质- rna相互作用酶联免疫吸附试验（PDI-ELISA或PRI-ELISA）的快速、灵敏的高通量筛选方法，用于鉴定靶向蛋白质-核酸相互作用的化合物。我们以哺乳动物高迁移群蛋白AT-hook 2 （HMGA2）为目标蛋白，netropsin为HMGA2 - dna相互作用的抑制剂，验证了PDI-ELISA方法。通过这种方法，我们成功地从29种dna结合化合物中鉴定出几种HMGA2-DNA相互作用的抑制剂和激活剂。在这项筛选工作的指导下，我们发现netropsin， HMGA2 - DNA相互作用的特异性抑制剂，强烈抑制小鼠前脂肪细胞3T3-L1细胞向脂肪细胞的分化，最可能的抑制机制是通过阻止HMGA2与目标DNA序列的结合。该方法应广泛适用于鉴定调节许多dna结合或rna结合蛋白的化合物或蛋白质。
DNA-binding and RNA-binding proteins are usually considered ‘undruggable’ partly due to the lack of an efficient method to identify inhibitors from existing small molecule repositories. Here we report a rapid and sensitive high-throughput screening approach to identify compounds targeting protein–nucleic acids interactions based on protein–DNA or protein–RNA interaction enzyme-linked immunosorbent assays (PDI-ELISA or PRI-ELISA). We validated the PDI-ELISA method using the mammalian high-mobility-group protein AT-hook 2 (HMGA2) as the protein of interest and netropsin as the inhibitor of HMGA2–DNA interactions. With this method we successfully identified several inhibitors and an activator for HMGA2–DNA interactions from a collection of 29 DNA-binding compounds. Guided by this screening excise, we showed that netropsin, the specific inhibitor of HMGA2–DNA interactions, strongly inhibited the differentiation of the mouse pre-adipocyte 3T3-L1 cells into adipocytes, most likely through a mechanism by which the inhibition is through preventing the binding of HMGA2 to the target DNA sequences. This method should be broadly applicable to identify compounds or proteins modulating many DNA-binding or RNA-binding proteins.
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影响因子：
7.2
作者：
O'Donnell M;Langston L;Stillman B
通讯作者：
Stillman B
影响因子：
3
作者：
Makley LN;Gestwicki JE
通讯作者：
Gestwicki JE
影响因子：
14.9
作者：
HIBMA, MH;ELY, SJ;CRAWFORD, L
通讯作者：
CRAWFORD, L
DOI：
10.1016/s1357-2725(98)00016-8
发表时间：
1998-07-01
影响因子：
4
作者：
Goodwin, G
通讯作者：
Goodwin, G
影响因子：
30.8
作者：
Horikoshi, Momoko;Yaghootkar, Hanieh;Mook-Kanamori, Dennis O.;Sovio, Ulla;Taal, H. Rob;Hennig, Branwen J.;Bradfield, Jonathan P.;St Pourcain, Beate;Evans, David M.;Charoen, Pimphen;Kaakinen, Marika;Cousminer, Diana L.;Lehtimaki, Terho;Kreiner-Moller, Eskil;Warrington, Nicole M.;Bustamante, Mariona;Feenstra, Bjarke;Berry, Diane J.;Thiering, Elisabeth;Pfab, Thiemo;Barton, Sheila J.;Shields, Beverley M.;Kerkhof, Marjan;van Leeuwen, Elisabeth M.;Fulford, Anthony J.;Kutalik, Zoltan;Zhao, Jing Hua;den Hoed, Marcel;Mahajan, Anubha;Lindi, Virpi;Goh, Liang-Kee;Hottenga, Jouke-Jan;Wu, Ying;Raitakari, Olli T.;Harder, Marie N.;Meirhaeghe, Aline;Ntalla, Ioanna;Salem, Rany M.;Jameson, Karen A.;Zhou, Kaixin;Monies, Dorota M.;Lagou, Vasiliki;Kirin, Mirna;Heikkinen, Jani;Adair, Linda S.;Alkuraya, Fowzan S.;Al-Odaib, Ali;Amouyel, Philippe;Andersson, Ehm Astrid;Bennett, Amanda J.;Blakemore, Alexandra I. F.;Buxton, Jessica L.;Dallongeville, Jean;Das, Shikta;de Geus, Eco J. C.;Estivill, Xavier;Flexeder, Claudia;Froguel, Philippe;Geller, Frank;Godfrey, Keith M.;Gottrand, Frederic;Groves, Christopher J.;Hansen, Torben;Hirschhorn, Joel N.;Hofman, Albert;Hollegaard, Mads V.;Hougaard, David M.;Hyppoenen, Elina;Inskip, Hazel M.;Isaacs, Aaron;Jorgensen, Torben;Kanaka-Gantenbein, Christina;Kemp, John P.;Kiess, Wieland;Kilpelainen, Tuomas O.;Klopp, Norman;Knight, Bridget A.;Kuzawa, Christopher W.;McMahon, George;Newnham, John P.;Niinikoski, Harri;Oostra, Ben A.;Pedersen, Louise;Postma, Dirkje S.;Ring, Susan M.;Rivadeneira, Fernando;Robertson, Neil R.;Sebert, Sylvain;Simell, Olli;Slowinski, Torsten;Tiesler, Carla M. T.;Toenjes, Anke;Vaag, Allan;Viikari, Jorma S.;Vink, Jacqueline M.;Vissing, Nadja Hawwa;Wareham, Nicholas J.;Willemsen, Gonneke;Witte, Daniel R.;Zhang, Haitao;Zhao, Jianhua;Wilson, James F.;Stumvoll, Michael;Prentice, Andrew M.;Meyer, Brian F.;Pearson, Ewan R.;Boreham, Colin A. G.;Cooper, Cyrus;Gillman, Matthew W.;Dedoussis, George V.;Moreno, Luis A.;Pedersen, Oluf;Saarinen, Maiju;Mohlke, Karen L.;Boomsma, Dorret I.;Saw, Seang-Mei;Lakka, Timo A.;Koerner, Antje;Loos, Ruth J. F.;Ong, Ken K.;Vollenweider, Peter;van Duijn, Cornelia M.;Koppelman, Gerard H.;Hattersley, Andrew T.;Holloway, John W.;Hocher, Berthold;Heinrich, Joachim;Power, Chris;Melbye, Mads;Guxens, Monica;Pennell, Craig E.;Bonnelykke, Klaus;Bisgaard, Hans;Eriksson, Johan G.;Widen, Elisabeth;Hakonarson, Hakon;Uitterlinden, Andre G.;Pouta, Anneli;Lawlor, Debbie A.;Smith, George Davey;Frayling, Timothy M.;McCarthy, Mark I.;Grant, Struan F. A.;Jaddoe, Vincent W. V.;Jarvelin, Marjo-Riitta;Timpson, Nicholas J.;Prokopenko, Inga;Freathy, Rachel M.
通讯作者：
Freathy, Rachel M.