This Article Full Text (PDF) References Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via HighWire Citing Articles via ISI Web of Science (70) Citing Articles via Google Scholar Google Scholar Articles by Pantoliano, M. W. Articles by Salemme, F. R. Search for Related Content PubMed PubMed Citation Articles by Pantoliano, M. W. Articles by Salemme, F. R. Social Bookmarking                   What's this?

High-Density Miniaturized Thermal Shift Assays as a General Strategy for Drug Discovery

Millennium Pharmaceuticals Inc., Cambridge, MA

Eugene C. Petrella

3-Dimensional Pharmaceuticals, Inc., Exton, PA

Joseph D. Kwasnoski

3-Dimensional Pharmaceuticals, Inc., Exton, PA

Victor S. Lobanov

3-Dimensional Pharmaceuticals, Inc., Exton, PA

James Myslik

3-Dimensional Pharmaceuticals, Inc., Exton, PA

Edward Graf

3-Dimensional Pharmaceuticals, Inc., Exton, PA

Ted Carver

3-Dimensional Pharmaceuticals, Inc., Exton, PA

Eric Asel

3-Dimensional Pharmaceuticals, Inc., Exton, PA

Barry A. Springer

3-Dimensional Pharmaceuticals, Inc., Exton, PA

Pamela Lane

3-Dimensional Pharmaceuticals, Inc., Exton, PA

F. R. Salemme

3-Dimensional Pharmaceuticals, Inc., Exton, PA

More general and universally applicable drug discovery assay technologies are needed in order to keep pace with the recent advances in combinatorial chemistry and genomics-based target generation. Ligand-induced conformational stabilization of proteins is a well-understood phenomenon in which substrates, inhibitors, cofactors, and even other proteins provide enhanced stability to proteins on binding. This phenomenon is based on the energetic coupling of the ligand-binding and protein-melting reactions. In an attempt to harness these biophysical properties for drug discovery, fully automated instrumentation was designed and implemented to perform miniaturized fluorescence-based thermal shift assays in a microplate format for the high throughput screening of compound libraries. Validation of this process and instrumentation was achieved by investigating ligand binding to more than 100 protein targets. The general applicability of the thermal shift screening strategy was found to be an important advantage because it circumvents the need to design and retool new assays with each new therapeutic target. Moreover, the miniaturized thermal shift assay methodology does not require any prior knowledge of a therapeutic target's function, making it ideally suited for the quantitative high throughput drug screening and evaluation of targets derived from genomics.

CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				M. Koszelak-Rosenblum, A. C. Krol, D. M. Simmons, C. C. Goulah, L. Wroblewski, and M. G. Malkowski His-311 and Arg-559 Are Key Residues Involved in Fatty Acid Oxygenation in Pathogen-inducible Oxygenase J. Biol. Chem., September 5, 2008; 283(36): 24962 - 24971. [Abstract] [Full Text] [PDF]

				G. A. Senisterra, B. Soo Hong, H.-W. Park, and M. Vedadi Application of High-Throughput Isothermal Denaturation to Assess Protein Stability and Screen for Ligands J Biomol Screen, June 1, 2008; 13(5): 337 - 342. [Abstract] [PDF]

				C. Attali, C. Frolet, C. Durmort, J. Offant, T. Vernet, and A. M. Di Guilmi Streptococcus pneumoniae Choline-Binding Protein E Interaction with Plasminogen/Plasmin Stimulates Migration across the Extracellular Matrix Infect. Immun., February 1, 2008; 76(2): 466 - 476. [Abstract] [Full Text] [PDF]

				W. J. Metzler, J. Yanchunas, C. Weigelt, K. Kish, H. E. Klei, D. Xie, Y. Zhang, M. Corbett, J. K. Tamura, B. He, et al. Involvement of DPP-IV catalytic residues in enzyme-saxagliptin complex formation Protein Sci., February 1, 2008; 17(2): 240 - 250. [Abstract] [Full Text] [PDF]

				H.-S. Shieh, K. J. Mathis, J. M. Williams, R. L. Hills, J. F. Wiese, T. E. Benson, J. R. Kiefer, M. H. Marino, J. N. Carroll, J. W. Leone, et al. High Resolution Crystal Structure of the Catalytic Domain of ADAMTS-5 (Aggrecanase-2) J. Biol. Chem., January 18, 2008; 283(3): 1501 - 1507. [Abstract] [Full Text] [PDF]

				E. Z. Baum, S. M. Crespo-Carbone, A. Klinger, B. D. Foleno, I. Turchi, M. Macielag, and K. Bush A MurF Inhibitor That Disrupts Cell Wall Biosynthesis in Escherichia coli Antimicrob. Agents Chemother., December 1, 2007; 51(12): 4420 - 4426. [Abstract] [Full Text] [PDF]

				G. L. Lorca, A. Ezersky, V. V. Lunin, J. R. Walker, S. Altamentova, E. Evdokimova, M. Vedadi, A. Bochkarev, and A. Savchenko Glyoxylate and Pyruvate Are Antagonistic Effectors of the Escherichia coli IclR Transcriptional Regulator J. Biol. Chem., June 1, 2007; 282(22): 16476 - 16491. [Abstract] [Full Text] [PDF]

				M. J. Baumann, J. M. Eklof, G. Michel, A. M. Kallas, T. T. Teeri, M. Czjzek, and H. Brumer III Structural Evidence for the Evolution of Xyloglucanase Activity from Xyloglucan Endo-Transglycosylases: Biological Implications for Cell Wall Metabolism PLANT CELL, June 1, 2007; 19(6): 1947 - 1963. [Abstract] [Full Text] [PDF]

				J. Ren, S. Sainsbury, S. E. Combs, R. G. Capper, P. W. Jordan, N. S. Berrow, D. K. Stammers, N. J. Saunders, and R. J. Owens The Structure and Transcriptional Analysis of a Global Regulator from Neisseria meningitidis J. Biol. Chem., May 11, 2007; 282(19): 14655 - 14664. [Abstract] [Full Text] [PDF]

				T. M. Mezzasalma, J. K. Kranz, W. Chan, G. T. Struble, C. Schalk-Hihi, I. C. Deckman, B. A. Springer, and M. J. Todd Enhancing Recombinant Protein Quality and Yield by Protein Stability Profiling J Biomol Screen, April 1, 2007; 12(3): 418 - 428. [Abstract] [PDF]

				C. Schalk-Hihi, H.-C. Ma, G. T. Struble, S. Bayoumy, R. Williams, E. Devine, I. P. Petrounia, T. Mezzasalma, L. Zeng, C. Schubert, et al. Protein Engineering of the Colony-stimulating Factor-1 Receptor Kinase Domain for Structural Studies J. Biol. Chem., February 9, 2007; 282(6): 4085 - 4093. [Abstract] [Full Text] [PDF]

				C. Schubert, C. Schalk-Hihi, G. T. Struble, H.-C. Ma, I. P. Petrounia, B. Brandt, I. C. Deckman, R. J. Patch, M. R. Player, J. C. Spurlino, et al. Crystal Structure of the Tyrosine Kinase Domain of Colony-stimulating Factor-1 Receptor (cFMS) in Complex with Two Inhibitors J. Biol. Chem., February 9, 2007; 282(6): 4094 - 4101. [Abstract] [Full Text] [PDF]

				G. A. Senisterra, E. Markin, K. Yamazaki, R. Hui, M. Vedadi, and D. E. Awrey Screening for Ligands Using a Generic and High-Throughput Light-Scattering-Based Assay J Biomol Screen, December 1, 2006; 11(8): 940 - 948. [Abstract] [PDF]

				M. Vedadi, F. H. Niesen, A. Allali-Hassani, O. Y. Fedorov, P. J. Finerty Jr., G. A. Wasney, R. Yeung, C. Arrowsmith, L. J. Ball, H. Berglund, et al. Chemical screening methods to identify ligands that promote protein stability, protein crystallization, and structure determination PNAS, October 24, 2006; 103(43): 15835 - 15840. [Abstract] [Full Text] [PDF]

				M. D. Cummings, M. A. Farnum, and M. I. Nelen Universal Screening Methods and Applications of ThermoFluor(R) J Biomol Screen, October 1, 2006; 11(7): 854 - 863. [Abstract] [PDF]

				K. M. Comess, M. E. Schurdak, M. J. Voorbach, M. Coen, J. D. Trumbull, H. Yang, L. Gao, H. Tang, X. Cheng, C. G. Lerner, et al. An Ultraefficient Affinity-Based High-Throughout Screening Process: Application to Bacterial Cell Wall Biosynthesis Enzyme MurF J Biomol Screen, October 1, 2006; 11(7): 743 - 754. [Abstract] [PDF]

				A. Magnusdottir, P. Stenmark, S. Flodin, T. Nyman, M. Hammarstrom, M. Ehn, M A. Bakali H, H. Berglund, and P. Nordlund The Crystal Structure of a Human PP2A Phosphatase Activator Reveals a Novel Fold and Highly Conserved Cleft Implicated in Protein-Protein Interactions J. Biol. Chem., August 11, 2006; 281(32): 22434 - 22438. [Abstract] [Full Text] [PDF]

				H. K. Koblish, S. Zhao, C. F. Franks, R. R. Donatelli, R. M. Tominovich, L. V. LaFrance, K. A. Leonard, J. M. Gushue, D. J. Parks, R. R. Calvo, et al. Benzodiazepinedione inhibitors of the Hdm2:p53 complex suppress human tumor cell proliferation in vitro and sensitize tumors to doxorubicin in vivo Mol. Cancer Ther., January 1, 2006; 5(1): 160 - 169. [Abstract] [Full Text] [PDF]

				J. Yanchunas Jr., D. R. Langley, L. Tao, R. E. Rose, J. Friborg, R. J. Colonno, and M. L. Doyle Molecular Basis for Increased Susceptibility of Isolates with Atazanavir Resistance-Conferring Substitution I50L to Other Protease Inhibitors Antimicrob. Agents Chemother., September 1, 2005; 49(9): 3825 - 3832. [Abstract] [Full Text] [PDF]

				K. L. Constantine, S. R. Krystek, M. D. Healy, M. L. Doyle, N. O. Siemers, J. Thanassi, N. Yan, D. Xie, V. Goldfarb, J. Yanchunas, et al. Structural and functional characterization of CFE88: Evidence that a conserved and essential bacterial protein is a methyltransferase Protein Sci., June 1, 2005; 14(6): 1472 - 1484. [Abstract] [Full Text] [PDF]

				K. El Omari, L. E. Bird, C. E. Nichols, J. Ren, and D. K. Stammers Crystal Structure of CC3 (TIP30): IMPLICATIONS FOR ITS ROLE AS A TUMOR SUPPRESSOR J. Biol. Chem., May 6, 2005; 280(18): 18229 - 18236. [Abstract] [Full Text] [PDF]

				T. E. Carver, B. Bordeau, M. D. Cummings, E. C. Petrella, M. J. Pucci, L. E. Zawadzke, B. A. Dougherty, J. A. Tredup, J. W. Bryson, J. Yanchunas Jr., et al. Decrypting the Biochemical Function of an Essential Gene from Streptococcus pneumoniae Using ThermoFluor(R) Technology J. Biol. Chem., March 25, 2005; 280(12): 11704 - 11712. [Abstract] [Full Text] [PDF]

				M. T. Garzon, M. C. Lidon-Moya, F. N. Barrera, A. Prieto, J. Gomez, M. G. Mateu, and J. L. Neira The dimerization domain of the HIV-1 capsid protein binds a capsid protein-derived peptide: A biophysical characterization Protein Sci., June 1, 2004; 13(6): 1512 - 1523. [Abstract] [Full Text] [PDF]