Abstract
Rho GTPases have been implicated in diverse cellular functions and are potential therapeutic targets in inflammation, cancer, and neurologic diseases. Virtual screening of compounds that fit into surface grooves of RhoA known to be critical for guanine nucleotide exchange factor (GEF) interaction produced chemical candidates with minimized docking energy. Subsequent screening for inhibitory activity of RhoA binding to the Rho-GEF, LARG, identified a Rho-specific inhibitor as a lead compound capable of blocking RhoA–LARG interaction and RhoA activation by LARG specifically and dose dependently. A microscale thermophoresis analysis was applied to directly quantify the binding interaction of the lead inhibitor with RhoA target. The lead inhibitor highlights the principle that rational targeting of subfamily members of Rho GTPases is feasible and potentially useful in future drug design effort.
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References
Etienne-Manneville S, Hall A (2002) Rho GTPases in cell biology. Nature 420:629–635
Ridley AJ (2001) Rho family proteins: coordinating cell responses. Trends Cell Biol 11:471–477
Zohn IM, Campbell SL, Khosravi-Far R, Rossman KL, Der CL (1998) Rho family proteins and Ras transformation: the RHOad less traveled gets congested. Oncogene 17:1415–1438
Sahai E, Marshall CJ (2002) RHO-GTPases and cancer. Nat Rev Cancer 2:133–142
Boettner B, Van Aelst L (2002) The role of Rho GTPases in disease development. Gene 286:155–174
Nassar N, Cancelas J, Zheng J, Williams DA, Zheng Y (2006) Structure-function based design of small molecule inhibitors targeting Rho family GTPases. Curr Top Med Chem 6:1109–1116
Marchioni F, Zheng Y (2009) Targeting rho GTPases by peptidic structures. Curr Pharm Des 15:2481–2487
Sebti SM, Der CJ (2003) Opinion: searching for the elusive targets of farnesyltransferase inhibitors. Nat Rev Cancer 3:945–951
Genth H, Dreger SC, Huelsenbeck J, Just I (2008) Clostridium difficile toxins: more than mere inhibitors of Rho proteins. Int J Biochem Cell Biol 40:592–597
Gao Y, Dickerson JB, Guo F, Zheng J, Zheng Y (2004) Rational design and characterization of a Rac GTPase-specific small molecule inhibitor. Proc Natl Acad Sci USA 101:7618–7623
Onesto C, Shutes A, Picard V, Schweighoffer F, Der CJ (2008) Characterization of EHT 1864, a novel small molecule inhibitor of Rac family small GTPases. Methods Enzymol 439:111–129
Narumiya S, Ishizaki T, Uehata M (2000) Use and properties of ROCK-specific inhibitor Y-27632. Methods Enzymol 325:273–284
Evelyn CR et al (2010) Design, synthesis and prostate cancer cell-based studies of analogs of the Rho/MKL1 transcriptional pathway inhibitor, CCG-1423. Bioorg Med Chem Lett 20:665–672
Vetter IR, Wittinghofer A (2001) The guanine nucleotide-binding switch in three dimensions. Science 294:1299–1304
Rossman KL, Der CJ, Sondek J (2005) GEF means go: turning on RHO GTPases with guanine nucleotide-exchange factors. Nat Rev Mol Cell Biol 6:167–180
Kristelly R, Gao G, Tesmer JJ (2004) Structural determinants of RhoA binding and nucleotide exchange in leukemia-associated Rho guanine-nucleotide exchange factor. J Biol Chem 279:47352–47362
Krieger E, Nielsen JE, Spronk CA, Kollman PA (2006) Fast empirical pKa prediction by Ewald summation. J Mol Graph Model 25:481–486
Krieger E, Darden T, Nabuurs S, Finkelstein A, Vriend G (2004) Making optimal use of empirical energy functions: force-field parameterization in crystal space. Proteins 57:678–683
Wienken CJ, Baaske P, Rothbauer U, Braun D, Duhr S (2010) Protein-binding assays in biological liquids using microscale thermophoresis. Nat Commun 19:100
Fukuhara S, Chikumi H, Gutkind JS (2000) Leukemia-associated Rho guanine nucleotide exchange factor (LARG) links heterotrimeric G proteins of the G(12) family to Rho. FEBS Lett 485:183–188
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Shang, X., Zheng, Y. (2012). Rational Design of Rho GTPase-Targeting Inhibitors. In: Zheng, Y. (eds) Rational Drug Design. Methods in Molecular Biology, vol 928. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-008-3_3
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DOI: https://doi.org/10.1007/978-1-62703-008-3_3
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