Rapid measurement of binding constants and heats of binding using a new titration calorimeter.
TL;DR: A new titration calorimeter is described and results are presented for the binding of cytidine 2'-monophosphate (2'CMP) to the active site of ribonuclease A.
About: This article is published in Analytical Biochemistry.The article was published on 1989-05-15. It has received 2561 citations till now. The article focuses on the topics: Binding constant & Isothermal titration calorimetry.
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TL;DR: A cell-permeable small molecule (JQ1) that binds competitively to acetyl-lysine recognition motifs, or bromodomains is reported, establishing proof-of-concept for targeting protein–protein interactions of epigenetic ‘readers’, and providing a versatile chemical scaffold for the development of chemical probes more broadly throughout the b romodomain family.
Abstract: Epigenetic proteins are intently pursued targets in ligand discovery. So far, successful efforts have been limited to chromatin modifying enzymes, or so-called epigenetic 'writers' and 'erasers'. Potent inhibitors of histone binding modules have not yet been described. Here we report a cell-permeable small molecule (JQ1) that binds competitively to acetyl-lysine recognition motifs, or bromodomains. High potency and specificity towards a subset of human bromodomains is explained by co-crystal structures with bromodomain and extra-terminal (BET) family member BRD4, revealing excellent shape complementarity with the acetyl-lysine binding cavity. Recurrent translocation of BRD4 is observed in a genetically-defined, incurable subtype of human squamous carcinoma. Competitive binding by JQ1 displaces the BRD4 fusion oncoprotein from chromatin, prompting squamous differentiation and specific antiproliferative effects in BRD4-dependent cell lines and patient-derived xenograft models. These data establish proof-of-concept for targeting protein-protein interactions of epigenetic 'readers', and provide a versatile chemical scaffold for the development of chemical probes more broadly throughout the bromodomain family.
3,489 citations
2,804 citations
1,648 citations
TL;DR: Crystal structures of complexes between the N-terminal domain of the yeast Hsp90 chaperone and ADP/ATP unambiguously identify a specific adenine nucleotide binding site homologous to the ATP-binding site of DNA gyrase B, suggesting that geldanamycin acts by blocking the binding of nucleotides to Hsp 90 and not the binding to incompletely folded client polypeptides as previously suggested.
1,258 citations
Cites methods from "Rapid measurement of binding consta..."
...Physical interaction ofdescribed elsewhere (Wiseman et al., 1989; Ladbury and Chowdhry, mammalian CDC37 with CDK4....
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TL;DR: The adaptor protein Hop mediates the association of the molecular chaperones Hsp70 and Hsp90 with the help of the TPR domains, which participate in the ordered assembly of HSp70-Hsp90 multichaperone complexes.
1,173 citations
Cites methods from "Rapid measurement of binding consta..."
...The position Binding of protein fragments and peptides to the TPR domains was measured by isothermal titration calorimetry (Wiseman et al., 1989) of one nickel atom was determined by analysis of an anomalous...
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...The position Binding of protein fragments and peptides to the TPR domains was measured by isothermal titration calorimetry (Wiseman et al., 1989)of one nickel atom was determined by analysis of an anomalous using a MicroCal MCS titration calorimeter (MicroCal Inc., North- domain of Hsc70 provides…...
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Book•
01 Jun 1969
TL;DR: In this paper, Monte Carlo techniques are used to fit dependent and independent variables least squares fit to a polynomial least-squares fit to an arbitrary function fitting composite peaks direct application of the maximum likelihood.
Abstract: Uncertainties in measurements probability distributions error analysis estimates of means and errors Monte Carlo techniques dependent and independent variables least-squares fit to a polynomial least-squares fit to an arbitrary function fitting composite peaks direct application of the maximum likelihood. Appendices: numerical methods matrices graphs and tables histograms and graphs computer routines in Pascal.
12,737 citations
TL;DR: Numerical methods matrices graphs and tables histograms and graphs computer routines in Pascal and Monte Carlo techniques dependent and independent variables least-squares fit to a polynomial least-square fit to an arbitrary function fitting composite peaks direct application of the maximum likelihood.
Abstract: Uncertainties in measurements probability distributions error analysis estimates of means and errors Monte Carlo techniques dependent and independent variables least-squares fit to a polynomial least-squares fit to an arbitrary function fitting composite peaks direct application of the maximum likelihood. Appendices: numerical methods matrices graphs and tables histograms and graphs computer routines in Pascal.
10,546 citations
TL;DR: The chapter discusses the stability of proteins and presents the results obtained on small compact globular proteins, which represent one single cooperative system, and the temperature-induced changes in protein, denaturational and predenaturational changes inprotein, thermodynamics of protein unfolding, and thermodynamic properties of protein.
Abstract: Publisher Summary The chapter discusses the stability of proteins and presents the results obtained on small compact globular proteins, which represent one single cooperative system. Protein is a cooperative system and behaves in an all-or-none fashion. Sharp changes in the properties of a protein do not mean anything in themselves because sequential multistep transitions exhibit the same sharp sigmoidal changes in the observed parameters. The problem of stability of native proteins is closely connected with the problem of protein denaturation, as stability can be judged only by breaking the native structure—that is, denaturing protein by various treatments. The pH of the solution is one of the most important factors determining the state of a protein. Potentiometric titration of protein revealed that smooth changes are connected with the titration of groups with a pK not very different from that of free amino acids, while the gross conformational changes associated with pH denaturation are accompanied by the unmasking of buried groups. The chapter also discusses the temperature-induced changes in protein, denaturational and predenaturational changes in protein, thermodynamics of protein unfolding, and thermodynamic properties of protein.
2,037 citations
TL;DR: This chapter focuses on the stability of protein structure and hydrophobic interaction, and examines the main achievements of microcalorimetric studies of protein denaturation and of the dissolution of nonpolar substances in water.
Abstract: Publisher Summary This chapter focuses on the stability of protein structure and hydrophobic interaction The interest in hydrophobic interactions was stimulated by their unusual thermodynamic properties: it is believed that they are governed, not by enthalpic, but by entropic features, characterized by the undesirable entropy decrease of water in the vicinity of nonpolar groups The amount of polar groups in proteins is almost the same as the amount of nonpolar ones; and according to crystallographic studies, most of them are arranged at distances suggesting hydrogen bond formation Hydrogen bonds were invoked to various degrees of importance in explaining the stabilization of the native structure The chapter examines the main achievements of microcalorimetric studies of protein denaturation and of the dissolution of nonpolar substances in water The chapter also discusses calorimetric studies of protein denaturation The denaturational increment of the heat capacity can be partly explained by a gradual melting of the residual structure in the denatured protein on heating The large negative entropy of the transfer of a nonpolar substance to water at room temperature indicates a definite increase of the order in water in the presence of such solutes Of the two approaches to decomposing the thermodynamics for dissolution of nonpolar solutes into water, the first, from a reference point at the maximum of the free energy of transfer, leads to the concept of the compact state of the nonpolar substance
1,231 citations