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Biomedical subjects

Eric V Anslyn

Publications and source records attributed to Eric V Anslyn.

At least 37 records · Page 2Linked to original sources
PubMed2004

Catalytic signal amplification using a Heck reaction. An example in the fluorescence sensing of CuII.

Catalytic signal enhancement using an organometallic reaction is demonstrated. The reactivity of a Heck cross-coupling reaction that creates a fluorophore is modulated by the addition of a polyazacyclam inhibitor. The inhibitor will complex with Cu(II), which restores the activity of the Pd(II). The addition of Cu(II) therefore leads to the generation of fluorescence, thereby creating a very sensitive assay for Cu(II). The rate of the Heck reaction is followed by monitoring emission as a function of time. The rate is proportional to the Cu(II) concentration and correlates to the affinity of the inhibitor to various metals. This strategy represents a general technique that can be exploited with other catalytic organometallic reactions.

Aza Compounds↗
PubMed2004

Guanidinium groups act as general-acid catalysts in phosphoryl transfer reactions: a two-proton inventory on a model system.

Cleavage/transesterification of phosphodiesters is catalyzed by various acidic groups in solution and with enzymes. General-acid catalysts can transfer protons to the developing phosphorane intermediate, resulting in a monoprotic-monoanionic intermediate, giving the so-called "triester mechanism". Using a proton inventory on a model compound (1) possessing an intramolecular hydrogen bond between a phosphodiester and a guanidinium group, we find that two protons move in the rate-determining step for cleavage/transesterification. In contrast, HPNP shows a single-proton inventory and is a substrate well accepted to react with the movement of only one proton at the transition state. We therefore propose a mechanism for 1 that involves general-acid catalysis by the guanidinium group. This leads one to conclude that other, more acidic groups, such as ammonium and imidazolium, would also act as general-acid catalysts.

Catalysis↗
PubMed2004

Thermodynamic analysis of receptors based on guanidinium/boronic acid groups for the complexation of carboxylates, alpha-hydroxycarboxylates, and diols: driving force for binding and cooperativity.

The thermodynamics of guanidinium and boronic acid interactions with carboxylates, alpha-hydroxycarboxylates, and diols were studied by determination of the binding constants of a variety of different guests to four different hosts (7-10). Each host contains a different combination of guanidinium groups and boronic acids. The guests included molecules with carboxylate and/or diol moieties, such as citrate, tartrate, and fructose, among others. The Gibbs free energies of binding were determined by UV/Vis absorption spectroscopy, by use of indicator displacement assays. The receptor based on three guanidinium groups (7) was selective for the tricarboxylate guest. The receptors that incorporated boronic acids (8-10) had higher affinities for guests that included alpha-hydroxycarboxylate and catechol moieties over guests containing only carboxylates or alkanediols. Isothermal titration calorimetry revealed the enthalpic and entropic contributions to the Gibbs free energies of binding. The binding of citrate and tartrate was investigated with hosts 7-10, for which all the binding events were exothermic, with positive entropy. Because of the selectivity of hosts 8-10, a simple boronic acid (14) was also investigated and determined to be selective for alpha-hydroxycarboxylates and catechols over amino acids and alkanediols. Further, the cooperativity of 8 and 9 in binding tartrate was also investigated, revealing little or no cooperativity with 8, but negative cooperativity with 9. A linear entropy/enthalpy compensation relationship for all the hosts 7-10, 14, and the carboxylate-/diol-containing guests was also obtained. This relationship indicates that increasing enthalpy of binding is offset by similar losses in entropy for molecular recognition involving guanidinium and boronic acid groups.

Binding Sites↗
PubMed2004

Threshold detection using indicator-displacement assays: an application in the analysis of malate in Pinot Noir grapes.

The mathematics for modeling indicator-displacement assay isotherms is presented and contrasted to the classical host-guest binding isotherm. It is shown that the signal response can be tuned to occur closer to 1 equiv of guest relative to a standard binding algorithm. This delay in response leads to a better triggering protocol for threshold detection schemes. The determination of malate in Pinot Noir must was calculated using this new mathematical model, which demonstrates how a color change can be tuned to occur near a desired concentration of analyte.

Algorithms↗
PubMed2004

Cooperative metal-coordination and ion pairing in tripeptide recognition.

[structure: see text] The recognition of tripeptides by a metal-centered receptor consisting of a rigid backbone region and variable tripeptide arms is described. The studies showed that the receptor was selective for l-Xxx-l-Lys-l-Lys, with Xxx = His, Cys, and Met, giving association constants near 1.0 x 10(6) M(-)(1). Binding enhancement through cooperative interactions of the peptidic arms is demonstrated.

Amino Acids↗
PubMed2004

Facile quantification of enantiomeric excess and concentration with indicator-displacement assays: an example in the analyses of alpha-hydroxyacids.

An enantioselective indicator-displacement assay was developed to determine enantiomeric excess (ee) and concentration of alpha-hydroxyacid samples with two independent absorption measurements. The inherent relationship between the absorbance of the indicator-displacement ensemble and the overall concentration and ee of the analyte was established through solution equilibria.

Catalysis↗
PubMed2004

Rate of enolate formation is not very sensitive to the hydrogen bonding ability of donors to carboxyl oxygen lone pair acceptors; a ramification of the principle of non-perfect synchronization for general-base-catalyzed enolate formation.

Two series of structures (1 and 2) possessing intramolecular hydrogen bonds to the lone-pair electrons of carbonyl oxygens have been examined to reveal the influence of the pK(a) of the hydrogen-bond donor on the rate of general-base-catalyzed enolate formation. The geometry of the hydrogen bonds is well accepted to be appropriate for intramolecular hydrogen-bond formation. Yet, as revealed by Brønsted plots, both series show very little dependence of the rate of enolate formation on the hydrogen-bond donor ability. The intramolecular hydrogen bonds give rate enhancements only on the order of 10-100-fold, and corrected Brønsted alpha-values are slightly below 0.1. The results can be understood by interpreting them in light of the Principle of Non-Perfect Synchronization. The results are consistent with the proton transfer occurring through an asynchronous transition state with the developing negative charge localized on carbon. We postulate that catalysts of enolate formation will be most effective if the binding groups are focused on stabilizing negative charge that is forming on the enolate carbon rather than on the enolate oxygen.

Alcohols↗
PubMed2004

A far-red fluorescent contrast agent to image epidermal growth factor receptor expression.

Recent developments in optical technologies have the potential to improve the speed and accuracy of screening and diagnosis of curable precancerous lesions and early cancer, thereby decreasing the costs of detection and management of epithelial malignancies. The development of molecular-specific contrast agents for markers of early neoplastic transformation could improve the detection and molecular characterization of premalignant lesions. In the oral cavity, epidermal growth factor receptor (EGFR) overexpression has been identified in early stages of premalignant lesions of the oral squamous cell carcinoma; therefore, real-time assessment of EGFR expression could serve as a biomarker for oral neoplasia. The purpose of our study was to develop a molecular-specific optical contrast agent targeted against EGFR for in vivo assessment of epithelial neoplasia using a monoclonal antibody and the far-red fluorescent dye, Alexa Fluor 660 streptavidin. In addition to demonstrating the specificity of the contrast agent for EGFR in cell lines, we document the ability to achieve penetration through 500 microm thick epithelial layers using multilayer tissue constructs and permeability-enhancing agents. Finally, using the fluorescence intensity of the contrast agent on fresh oral cavity tissue sections, we were able to distinguish abnormal from normal oral tissue. This contrast agent should have important clinical applications for use in conjunction with fluorescence spectroscopy or imaging (or both) to facilitate tumor detection and demarcation.

Biopsy↗
PubMed2003

2,6-di(pyrimidin-4-yl)pyridine ligands with nitrogen-containing auxiliaries: the formation of functionalized molecular clefts upon metal coordination.

A series of ligands (1-4) based on a 2,6-di(pyrimidin-4-yl)pyridine scaffold have been synthesized, and their abilities to form complexes with Zn(II) and Cu(II) have been determined using UV/vis spectroscopy in buffered aqueous solution (0.01 M N-[2-hydroxyethyl]piperazine-N'-[3-ethanesulfonic acid] (HEPES) at pH = 6.8). The Zn(II) complex of 1 was determined to have a formation constant of 8.4 x 10(3) M(-)(1) while the formation constant of the Cu(II) complex was found to be 1 x 10(6) M(-)(1). The presence of auxiliary amines in 2 increased the stability of the Zn(II) complex relative to that of 1 by a factor of over 40, suggesting possible coordination of the auxiliaries to the Zn(II) center. The guanidinium and 2-amino-4,5-dihydro-imidazolinium groups of 3 and 4 considerably diminished the stability of the Zn(II) and Cu(II) complexes relative to those of 1. X-ray crystal structures of 1-Zn, 3-Zn, 4, and 4-Zn were obtained and are discussed. A significant increase in the stability of 3-Zn, but not in the stability 1-Zn, was observed upon the addition of 1 equiv of sodium phosphate, implicating a stabilizing interaction of the guanidinium groups of 3-Zn and the phosphate anion.

Cations, Divalent↗
PubMed2003

Triton X-100 enhances ion-pair-driven molecular recognition in aqueous media. Further work on a chemosensor for inositol trisphosphate.

Molecular recognition in water based on ion-pairing is hampered by competition from the solvent. We have reported that synthetic receptor 1 recognizes inositol trisphosphate (IP3) with a binding constant of 10(8) M(-1) in methanol. In this paper, instead of working in methanol, we report that the surfactant Triton-X 100 also yields a large absorbance or fluorescence change and a binding constant on the order of 10(6) M(-1).

Biosensing Techniques↗
PubMed2003

Energetics of phosphate binding to ammonium and guanidinium containing metallo-receptors in water.

The design and synthesis of receptors containing a Cu(II) binding site with appended ammonium groups (1) and guanidinium groups (2), along with thermodynamics analyses of anion binding, are reported. Both receptors 1 and 2 show high affinities (10(4) M(-1)) and selectivities for phosphate over other anions in 98:2 water:methanol at biological pH. The binding of the host-guest pairs is proposed to proceed through ion-pairing interactions between the charged functional groups on both the host and the guest. The affinities and selectivities for oxyanions were determined using UV/vis titration techniques. Additionally, thermodynamic investigations indicate that the 1:phosphate complex is primarily entropy driven, while the 2:phosphate complex displays both favorable enthalpy and entropy changes. The thermodynamic data for binding provide a picture of the roles of the host, guest, counterions, and solvent. The difference in the entropy and enthalpy driving forces for the ammonium and guanidinium containing hosts are postulated to derive primarily from differences in the solvation shell of these two groups.

Binding Sites↗
PubMed2003

Studies into the thermodynamic origin of negative cooperativity in ion-pairing molecular recognition.

The association of synthetic receptors to target guests often proceeds through the cooperative action of multiple binding forces. An investigation into the thermodynamic origin of cooperativity in ion-pairing host-guest binding in water is described. The binding affinities of 1,2,3,4-butanetetracarboxylate, tricarballate, glutarate, and acetate to a C(3)(v) symmetric metallo-host (1) are characterized in terms of the binding constants (K(a)) and the thermodynamic parameters deltaG degrees, deltaH degrees, and deltaS degrees, as determined by isothermal titration calorimetry (ITC). These values are used to determine the individual contributions of the binding interaction to the overall binding. Several ways to view the combination of the individual binding events that make up the whole are analyzed, all of which lead to the conclusion of negative cooperativity. Combined, the data were used to evaluate the thermodynamic origin of negative cooperativity for this series of guests, revealing that entropy is the largest contributing factor. An interpretation of this result focuses upon differences in the number of water molecules displaced upon binding.

Acetic Acid↗
PubMed2003

Determination of inorganic phosphate in serum and saliva using a synthetic receptor.

[structure: see text] A C(3)(v) symmetric synthetic receptor (1) was employed in an indicator-displacement assay to determine the phosphate concentrations in both horse serum and human saliva at biological pH. The determination of the phosphate concentrations in the serum and saliva using the colorimetric assay were 1.6 and 5.1 mM, respectively. These results further accentuate the usefulness of synthetic receptors in truly practical applications.

Animals↗
PubMed2003

C3v symmetric receptors show high selectivity and high affinity for phosphate.

The binding of phosphate to tripodal metalloreceptors 1 and 2 is reported. Receptors 1 and 2 are C3v symmetric, designed to complement three sides of a tetrahedron. The receptors derive from tripodal ligands that are preorganized through binding to a central Cu(II) atom. These metalloreceptors demonstrate high selectivity and affinity for the molecular recognition of phosphate in aqueous media at neutral pH. The binding of phosphate and other anions to the cavities of receptors 1 and 2 was monitored by UV/vis titration techniques. Binding algorithms were used to determine the affinity of phosphate to 1 and 2 with association values (Ka) of 2.5 x 104 and 1.5 x 104 M-1, respectively.

Anions↗
PubMed2003

Differential receptors create patterns diagnostic for ATP and GTP.

Herein we report the combination of a library of resin-bound sensors along with a multicomponent sensor array. This novel combinatorial array sensor system shows selectivity for nucleotide phosphates in solution. The design of the anchored receptor includes a 1,3,5-trisubstituted-2,4,6-triethylbenzene scaffold coupled with peptide libraries. Each chemosensor is placed into a micromachined cavity within a silicon wafer, and the optical changes observed by a charged-coupled device result in near-real-time digital analysis of solutions. A colorimetric displacement assay was performed, and time-dependent imaging studies of the selected sensing ensembles result in a differential responses upon addition of adenosine 5'-triphosphate (ATP), adenosine 5'-monophosphate (AMP), or guanosine 5'-triphosphate (GTP). An advantage to this approach is that it creates an array of sensors that gives a fingerprint response for each analyte. Principal component analysis indicates that the library of chemosensors can differentiate between ATP, GTP, and AMP. On the basis of factor loading values, individual sensors from the library were sequenced to elucidate their chemical composition.

Adenosine Triphosphate↗