Balancing interactions in proline-based receptors for chiral recognition of l-/d-DOPA.

Proline based receptors (1-14) attached with phenylboronic acid and benzaldehyde binding groups at the N-/C- or C-/N-termini of the proline residue were created for chiral recognition of l-/d-DOPA, in an attempt to examine if balancing the two binding events would influence the recognition. By changing the positions of boronic acid and aldehyde groups substituted on the phenyl rings (1-4, 5-8) and the site at which phenylboronic acid and benzaldehyde moieties attached respectively to the N- and C-termini or C- and N-termini of the proline residue (1-4vs.5-8), and by introducing an electron-withdrawing fluorine atom in the phenyl ring of the weaker binder the benzaldehyde moiety (11vs.1, 14vs.5), we were able to show that a better balance of the two binding events does improve the chiral recognition. This finding can only be made with the current version of receptors that were equipped with two different binding groups. Together with the finding that the chiral recognition performance in mixed organic-aqueous solutions is tunable by varying the solvent composition, we have now arrived at a protocol for designing proline based receptors for extended applications in chiral recognition.

Proline-Based Boronic Acid Receptors for Chiral Recognition of Glucose.

Chiral recognition remains a major challenge in the area of molecular receptor design. With this research, we set out to explore the use of proline-based receptors for chiral recognition. Importantly, the proline structure allows for the introduction of at least two different binding groups due to the availability of both an amine and carboxylic acid group. Here we report a proof-of-concept exploration into the chiral recognition of d/l-glucose as a model chiral species, which prefers to bind to at least two boronic acid groups. We evaluated several proline-based receptors incorporating two phenylboronic acid groups, respectively, at the N- and C-termini of the amino acid residue, via amide bonds. We confirmed that the receptors exhibited chiral recognition using CD, 1H NMR, and 19F NMR spectroscopy. Given the derivation diversity available, our strategy to use proline-based receptors for chiral recognition holds significant promise for extension to other chiral systems.

Aggregation-Switching Strategy for Promoting Fluorescent Sensing of Biologically Relevant Species: A Simple Near-Infrared Cyanine Dye Highly Sensitive and Selective for ATP.

We report a strategy for enhanced performance of fluorescent sensing of biologically relevant species that often bind with natural receptors via multiple interactions. We propose making a fluorescent sensory molecule to form H-aggregates such that its emission is quenched leaving a low background, and upon binding to a biologically relevant species, the aggregate switches to another form in which the fluorescent species is better protected to afford a stronger emission signal. Meanwhile, the aggregated fluorescent dyes afford multiple interactions with the sensing species that require multiple binding sites. The lower background, stronger binding, and stronger signal would therefore lead to a much higher sensing performance, as improved selectivity would also result in along with the signal amplification. We thus designed a near-IR cyanine dye bearing two boronic acid groups (Cy-BA) for fluorescent sensing of ATP such that the boronic acid groups in the dye molecule bind to the cis-diol moiety in ATP. Introduction of the cationic surfactant dodecyltrimethylammonium bromide (DTAB) below its critical aggregation concentration is key because Cy-BA molecules made into H-aggregates were practically nonfluorescent. Upon mixing with ATP, a dramatic enhancement in the fluorescence occurred because of the formation of ATP/Cy-BA/DTAB vesicles in which the fluorescent dye is well dispersed and protected. This sensing scheme, despite the dynamic nature of the boronic acid/cis-diol interaction, weakness of the electrostatic interactions among ATP/Cy-BA/DTAB, and poor selectivity of these interactions, allows for highly sensitive and selective detection of ATP in aqueous solution.

Iron(III)-Selective Chelation-Enhanced Fluorescence Sensing for In Vivo Imaging Applications.

A "turn-on" pattern Fe(3+) -selective fluorescent sensor was synthesized and characterized that showed high fluorescence discrimination of Fe(3+) over Fe(2+) and other tested ions. With a 62-fold fluorescence enhancement towards Fe(3+) , the probe was employed to detect Fe(3+) in vivo in HeLa cells and Caenorhabditis elegans, and it was also successfully used to elucidate Fe(3+) enrichment and exchange infected by innexin3 (Inx3) in hemichannel-closed Sf9 cells.

Highly selective in vivo imaging of endogenous/exogenous phosphate ion over ATP and PPi.

A chemodosimeter was designed to function as a highly selective phosphate ion (Pi ) sensor, showing a 91-fold ratiometric fluorescence enhancement. The probe successfully visualized exogenous and endogenous apyrase-catalyzed Pi generation and was the first probe able to trace the generation and enrichment of Pi through hemichannel closure in Sf9 cells.

Phosphate ion targeted colorimetric and fluorescent probe and its use to monitor endogeneous phosphate ion in a hemichannel-closed cell.

Fluorescent probe 1, the first inorganic phosphate (Pi) targeted colorimetric and fluorescent probe to detect endogenous Pi in hemichannel-closed cells, has been developed. Probe 1 undergoes a unique Pi induced hydrolytic reaction in DMSO-HEPES (V/V = 9:1) buffered (0.02 M, pH 7.4) solutions that produces a colorimetric change associated with a 62 nm red-shift in the UV-vis absorption maximum and up to a 780-fold enhancement in the fluorescence intensity. The mechanistic proposal that these spectroscopic changes are associated with reaction Pi with 1 to form coumarin gains support from the results of theoretical calculations and mass spectrometry studies. Observations made in fluorescence imaging studies with HeLa cells and C. elegans show that 1 can be employed to monitor Pi production in vivo caused by apyrase-catalyzed ATP hydrolysis. Moreover, probe 1 was utilized to show that apoptosis of hemichannel-closed Sf9 cells is caused by Inx3 promoted dephosphorylation of Akt (RAC serine/threonine-protein kinase), leading to an elevation of the concentration of Pi. Overall, the study has produced the first fluorescent sensor 1 for endogenous inorganic phosphate. Moreover, the utility of 1 for measuring Pi release in vitro has been demonstrated and utilized to elucidate the mechanism of Inx3 action in hemichannel-closed Sf9 cells.

A lysosome-targeted fluorescent chemodosimeter for monitoring endogenous and exogenous hydrogen sulfide by in vivo imaging.

A lysosome-targeted fluorescent chemodosimeter, 1, was developed for monitoring endogenous and exogenous H2S by in vivo imaging of HeLa cells, D. melanogaster and C. elegans. In the tests of mutated C. elegans (SRP-6 nulls), chemodosimeter 1 could trace the accumulation of lysosome and lysosomal injury with a high resolution.

A pyrene derivative for Hg(2+) -selective fluorescent sensing and its application in in vivo imaging.

An Hg(2+) -selective fluorescent sensor (1) bearing pyrene as a fluorophore was synthesized. A sandwich-stacking binding mode was formed during the binding process, which increased the excimer fluorescence 22-fold at 490 nm. Compound 1 was successfully applied in in vivo imaging to trace the enrichment and distribution of mercury in the nervous system, digestive system, and reproductive system of Caenorhabditis elegans, as well as the organs of zebrafish.