Search

This review covers the mechanistic origin of luminescent mono- and bimetallic gold-based nanomaterials and their syntheses by the chemical reduction of metal precursors in the presence of surface capping agents.

The key steps for seed mediated growth of noble metal nanoparticles involve primary and secondary nucleation, which depends upon the energy barrier and ligand supersaturation standards of the medium. Herein we report the unique case of methionine (Met) controlled growth reaction, which rather proceeds via impeding secondary nucleation in presence of citrate stabilized gold nanoparticle (AuNP). The interaction between freshly generated Au+ and thioether group of Met in the medium restricts the secondary nucleation process involving further Au+ reduction. This incomplete conversion of Au+ results in a significant enhancement of the zeta (ζ) potential even at low concentration of Met. Furthermore, the aurophilic interaction of Au+ controls the self-assembly process of the in situ generated emissive nucleated particles. Nucleation of primary particles on seed surface, their segregation and time dependent conversion to larger particles within self-assembly confirm the nonclassical growth, which has further been explored with Met containing bio-inspired peptides.

Time-dependent investigation of growth reactions from 0.45 nM gold nanoparticle seeds in the presence of 9 mM amino acids containing polar uncharged side chain (AApusc) serine (Ser), threonine (Thr...

Hydroxylamine-based growth reaction in the presence of natural l-amino acids (9 mM) and gold nanoparticle seed mostly produce aggregated or nonaggregated gold nanostructures except the cases of imm...

The formation of growth-mediated structures from gold nanoparticle seeds was studied in the presence of amine-modified single-strand DNA sequences and reducing agents such as hydroxylamine and hydroquinone. In the case of hydroxylamine, spherical gold nanoparticle seeds (0.45 nM) were incubated with amine-modified single-strand DNA probes PMR (amine-5'-ACATCAGT-3') and PML (amine-5'-GATAAGCT-3'), which resulted in gold nanoflowers and nanospheres, respectively. When the concentration of the nanoparticle seeds was varied (0.15-0.45 nM), only the PMR sequence showed growth-mediated development of gold nanoflowers. The size of the gold nanoparticles obtained is independent of the seed concentration for both PMR and PML sequences. In contrast, in the presence of the reducing agent hydroquinone, the growth processes are identical in for both the sequences. At a lower seed concentrations (0.15 nM), gold nanoflowers of larger size were observed for both sequences, whereas at higher seed concentrations (0.45 nM), much smaller gold nanospheres resulted. The formation and stability of nanoflowers and nanospheres for PMR and PML with hydroxylamine-based reduction were further studied in detail with diverse controlled amine-modified (5'-, 3'- and both end-modified) and non-modified DNA sequences with other mutants of these two sequences.

Addition of bis-pyrazole based cryptand regioisomers (1 and 2) to citrate stabilized gold nanoparticles (AuNPs) led to gold supraspheres (AuSSL) and elongated dodecahedron (AuEDL) supraparticles, respectively. These supraparticles as heterogenous hosts show a high uptake of a small fluorophore SEA-SC1 in an aqueous medium and reversible release in organic solvents.

The cellular delivery of oligonucleotides has been a major obstacle in the development of therapeutic antisense agents. PNAs (Peptide Nucleic Acid) are unique in providing a modular peptidic backbone that is amenable to structural and charge modulation. While cationic PNAs have been shown to be taken up by cells more efficiently than neutral PNAs, the generality of uptake across different nucleobase sequences has never been tested. Herein, we quantified the relative uptake of PNAs across a library of 10 000 sequences for two different PNA backbones (cationic and neutral) and identified sequences with high uptake and low uptake. We used the high uptake sequence as a bait for target identification, leading to the discovery that a protein, caprin‐1, binds to PNA with backbone and sequence discrimination. We further showed that purified caprin‐1 added to cell cultures enhanced the cellular uptake of PNA as well as DNA and RNA.

Hydroxylamine-based growth reaction in the presence of natural l-amino acids (9 mM) and gold nanoparticle seed mostly produce aggregated or nonaggregated gold nanostructures except the cases of immediate precipitation with aspartic acid, glutamic acid, cysteine, and tyrosine. Among the other amino acids, arginine shows the control growth reaction to form gold nanoflower from gold nanoparticle seeds, which were preincubated with amine-modified DNA (NH

We developed highly selective fluorescent Mg2+ probes based on a novel tridentate Mg2+-selective chelator. The superior selectivity for Mg2+ over Ca2+ enabled the detection of intracellular Mg2+ concentration changes without any response to Ca2+ influx. The new probes are potential keys to elucidating intracellular Mg2+ dynamics under conditions of extremely sharp Ca2+ fluctuation.

Nucleic acid templated reactions are enabled by the hybridization of probe-reagent conjugates resulting in high effective reagent concentration and fast chemical transformation. We have developed a reaction that harnesses cellular microRNA (miRNA) to yield the cleavage of a linker releasing fluorogenic rhodamine in a live vertebrate. The reaction is based on the catalytic photoreduction of an azide by a ruthenium complex. We showed that this system reports specific expression of miRNA in living tissues of a vertebrate., We have developed a reaction that harnesses cellular microRNA (miRNA) to yield the cleavage of a linker releasing fluorogenic rhodamine in a live vertebrate. The reaction is based on the catalytic photoreduction of an azide by a ruthenium complex.

In this study, we have compared neutral coumarin based well-known commercially available probes C6, C7 and C545T for fluorogenic response from the aggregated insulin. The immediate fluorogenic responses were comparatively poor from all the three probes with respect to the previously reported response from thioflavin T (ThT) in the presence of aggregated insulin. Interestingly C6 among the three neutral coumarin derivative showed a significant steady increase of fluorescence intensity with time up to 6 h before reaching the saturation limit. Similar time dependent fluorogenic experiment with C7, C545T and ThT showed comparatively fast saturation within few minutes to 2 h. The molecular docking and simulation studies showed that these neutral probes could be stabilized in the aggregated form of the insulin predominantly by non-covalent weak interactions such as hydrogen bonding, π-π and cation-π interactions. The probability distributions of the dihedral angles between two heterocyclic parts in C6 showed maximum probability of occurrence at 0° and 180°. These probability distributions of the dihedral angles between two heterocyclic parts within all the four fluorophores provided the justification of selective time dependent fluorescence enhancement from C6 in presence of insulin aggregate. The overall fluorogenic enhancement from C6 was comparable to the fluorogenic response from ThT and theoretical study confirmed distinctly different origin of this associated slow time dependent fluorogenic response.

Au–FexOy nanocomposites with a variable gold-to-iron ratio were stabilized with l-tryptophan. The synthetic methodology is based on the facile redox reaction between Au(III) and Fe(0) in the presence of gold nanoparticle as a seed at room temperature in an aqueous medium. The synthesis results in the deposition of Au nanoparticles on the surface of iron oxide layers. Composition variation in the nanocomposites was obtained by controlling the seed amount and reducing agent. These nanocomposites are used as electrocatalysts for the thermodynamically unfavorable oxygen evolution reaction (OER) from water. Among the nanocomposites, the most efficient OER activity was observed from the nanocomposite 12. The content of iron with respect to gold is at the maximum in the nanocomposite, which was obtained from the reaction with a minimum seed concentration and maximum reducing agent.

We developed highly selective fluorescent Mg

Controlled diluted aqua regia addition leads to NIR (near infrared) luminescence from aggregated gold nanoparticles at 916 nm. This turn-on luminescence has been observed for gold nanoparticles, regardless of reductants used in their preparation. These aggregated nanoparticles are nontoxic and have been used for bioimaging in human liver carcinoma cells.

A small molecule SEA-SC2 exhibited fluorescence in response to 10–60 nM amyloid beta (Aβ)40 and Aβ42 monomers at pH 7.4, in the presence of gold nanoparticle (AuNP). Here we report the fluorogenic detection due to the response from low nanomolar level of Aβ peptide monomer. The small organic molecule or AuNP has been reported to show detection limit up to 200 nM Aβ peptide. Furthermore, fluorescence enhancement studies with different stoichiometries of Aβ:Cu(II) confirmed the initial Aβ:Cu(II) adduct at the nanomolar level. This methodology has been applied to the detection of nanomolar concentrations of Aβ monomer in artificial cerebrospinal fluid and human serum albumin.

The programmability of oligonucleotide recognition offers an attractive platform for directing the assembly of ligands that can interact cooperatively with a target or the assembly of reactive partner that can engage in chemical reactions. In both cases, an oligonucleotide directs the assembly, which yields a functional output. In terms of the controlled display of ligands, several examples have shown a significant increase in binding upon ligand assembly using small molecule fragments, peptides, glycans, and protein fragments. Combinatorial approaches to identifying the optimal pairing have also been reported. In terms of nucleic acidtriggered reactions, several robust chemistries have been reported, leading to the unmasking of different fluorophores or bioactive molecules as well as the synthesis of a bioactive compound in response to DNA or even cellular RNA. Herein we present the historical context of this work and summarize recent developments in this area.

A metal ion sensing fluorophore L that exhibits a large two-photon absorption cross-section has been synthesized in good yields. The influences of different metal ion inputs, on the one- and two-photon spectroscopic properties of L , have been investigated. The ligand itself does not show any fluorescence although in presence of a metal ion like Zn(II), Cd(II), Mg(II) or Ca(II), a ∼25 time enhancement of fluorescence is observed. The ligand with symmetrical “donor–acceptor–acceptor–donor” characteristics exhibits a large two-photon absorption cross-section measured by femtosecond open-aperture Z-scan technique at 880 nm. However, presence of any of the above metal ions lowers its two-photon absorption cross-section ( δ ) to different extents at 880 nm. Theoretical calculation carried out in DFT formalism on the ligand and its Zn(II) complex corroborate experimental results.

A PET fluorescence signaling system L1 has been synthesized in the ‘Receptor1-Spacer-Fluorophore-Spacer-Receptor2’ format through attachment of a laterally non-symmetric heteroditopic cryptand and a 4,7,10,13-tetraoxa-1-azacyclopentadecane macrocycle at the 9- and 10-positions of anthracene. The cryptand receptor is derivatized with two 2,4-dinitrobenzene groups to impart structural rigidity. This system exhibits selective fluorescence enhancement in presence of Na(I) and Tl(I) ions simultaneously and mimics the AND logic function. The cryptand receptor also binds a Cu(II) ion outside the cavity and allows L1 to perform as an INHIBIT chemical logic in presence of H+ and Cu(II) ions.

Labelling of proteins with a luminescent ruthenium complex enables the direct visualization and photocatalytic reduction of aryl azide in live cells. The confinement of catalysis to the labeled proteins was visualized using an azide-based immolative linker releasing a precipitating dye.

Review: recent progress in the synthesis, screening, and self-assembly of nucleic acidtagged peptidic libraries

Cell permeable probes to image the presence and localization of kinases are important in studying their function and as diagnostic tools. Despite the central role of kinases as therapeutic targets, there are remarkably few probes available. Herein we report the discovery of a probe to image two therapeutically relevant kinases: EGFR and ERBB2. The probe was identified from a library based on a scaffold derived from the resorcyclic acid lactones that form a covalent adduct by reacting specifically with an unconserved cysteine in the nucleotide-binding site of the kinases. We demonstrated the utility of the newly discovered probe by imaging of EGFR localization and ERBB2 inhibition in live cells.

Reactions templated by cellular nucleic acids are attractive for nucleic acid sensing or responsive systems. Herein we report the use of a photocatalyzed reductive cleavage of an immolative linker to unmask a rhodamine fluorophore, and its application to miRNA imaging. The reaction was found to proceed with a very high turnover (>4000) and provided reliable detection down to 5 pm of template by using γ-serine-modified peptide nucleic acid (PNA) probes. The reaction was used for the selective detection of miR-21 in BT474 cells and miR-31 in HeLa cells following irradiation for 30 min. The probes were introduced by using reversible permeation with streptolysin-O (SLO) or a transfection technique.

The photoreduction of azide-based immolative linker by Ru(II) conjugates to uncage rhodamine was achieved using different oligomeric protein templates. The generality of the approach was validated with three sets of ligand having varying affinity to their target (biotin, desthiobiotin and raloxifene). The reaction rates of the templated reaction was found to be at least 30-fold faster than the untemplated reaction providing a clear fluorescent signal in response to the protein oligomer within 30 min. The templated reaction was found to also proceed in cellulo and could be used to identify acetyl coenzyme A carboxylase (ACC) in Pseudomonas aeruginosa and human cell lines as well the and estrogen receptor (ER).

The design and synthesis of molecular probes competent for pH signaling within or beyond a certain range is a complicated matter. Herein a new mechanism for ''OFF-ON-OFF'' absorbance and fluorescence intensities vs. pH behaviour is described. The probe design is based on the connection of carboxylic acid derivatized benzoxazole and 7-hydroxycoumarin/iminocoumarin parts. The protonation/deprotonation of the carboxylic acid (-COOH), N atom of benzoxazole ring and hydroxy part of the coumarin ring have been used for this mechanistic study. We have designed the molecule in such a fashion that deprotonation of the hydroxy part takes place at a lower pK(a) compared to deprotonation of the -COOH. The dual ''OFF-ON-OFF'' properties of our probes depend on the C-C bond between the two different heterocyclic parts. Quantum mechanical calculations showed that the particular 'C-C' bond has an additional π-character. The twisting around this bond in different forms is responsible for such an ''OFF-ON-OFF'' property. This mechanism is new in fluorescence alteration processes. The delocalization of charge from one heterocyclic part to the other heterocyclic part in the mono- and dianionic forms controls the ''OFF-ON-OFF'' properties. The role of the carboxylic acid group was examined using an acetyl substituted derivative. One of our probes was successfully applied in live cell imaging studies in media at different pH.

Two laterally non-symmetric aza-oxa cryptands have been derivatized with the electron-withdrawing fluorophore, 7-nitrobenz-2-oxa-1,3-diazole to obtain the corresponding mono-, bis- and tris-products. In each case, no appreciable emission is observed when the fluorophore is excited due to an efficient photoinduced intramolecular electron transfer (PET) from the lone pair on nitrogen present in the bridges. In the presence of a number of transition and heavy metal ions, their emission characteristics change. The electron-withdrawing ability of the fluorophore drastically alters the binding ability of the cryptand such that Hg(II) affords largest enhancement of fluorescence. In contrast, mono-, bis- and tris-anthryl (electron-donating) derivatives of these cryptands do not exhibit any selectivity towards Hg(II) and affords low fluorescence enhancement. Also, the difference in cavity dimension of the o-cryptand and m-cryptand plays a crucial role in terms of selectivity.

The targeted protein of interest is fused with genetically modified β-lactamase enzyme, which reacts with the probe in physiological conditions to break the aggregated interaction between the fluorophore and quencher. This alliance-separation technique is new for protein labeling and is probed in vitro and in live cell imaging studies.

A novel hybrid of azacrown ether-oxabridged fluorogenic chemosensor (1) has been synthesized: the metal ion binding ability of the hybrid molecule shows higher affinity towards transition metal ion Zn(2+) and heavy metal ion Hg(2+) compared to the parent azacrown ether which shows poor metal ion selectivity: the binding modes of the Zn(2+) and Na(+) towards have been discussed.

Two azine molecules, [3-{4-(dimethylamino)phenyl}prop-2-en-1-ylidene]hydrazone (HL) and [4-(dimethylamino)benzylidene]hydrazone (LL) synthesized via Schiff base condensation, show an absorption band due to intra-ligand charge transfer (ILCT). Both ligands show weak emission in the absence of a metal ion as input. When Ag(+) ions are added to either compound in THF, the metal ion gets bonded to the azine moiety resulting in a high intensity emission (approximately 400 fold). While Cu(+) shows a slight enhancement of emission, other first-row transition, alkali or alkaline-earth metal ions do not show any emission allowing Ag(+) ions to be detected in the presence of these metal ions. Both the Ag(+) complexes were characterised by X-ray crystallography and show solid-state spectra similar to their solution spectra. Time-resolved fluorescence measurements done on the complexes show two excited-state lifetimes.

The movement of a copper ion inside the cavity of cryptands having two distinct metal binding sites at each end is linked to a fluorescence ON-OFF process in a reversible manner, providing a new way of fluorescence switching.

A laterally nonsymmetric aza cryptand has been derivatized with one 7-nitrobenz-2-oxa-1,3-diazole (fluorophore(1)) and one/two anthracenes (fluorophore(2)) to obtain 1 and 2. Their emission characteristics are probed in the presence of a number of transition metals and proton. In the case of 1, Cu(II), Zn(II), Cd(II), and proton afford a large enhancement of fluorescence, whereas Fe(II) and Ag(I) exhibit one order of magnitude less enhancement. In contrast, 2 gives a large enhancement with Cu(II), Ag(I), and proton. The enhancement is observed in the diazole moiety even when the anthracene fluorophore is excited because of substantial fluorescence resonance energy transfer from anthracene to the diazole moiety. Compounds 1 and 2 can be termed as the second-generation fluorescence signaling systems.

Development of protein labeling techniques with small molecules is enthralling because this method brings promises for triumph over the limitations of fluorescent proteins in live cell imaging. This technology deals with the functionalization of proteins with small molecules and is anticipated to facilitate the expansion of various protein assay methods. A new straightforward aggregation and elimination-based technique for a protein labeling system has been developed with a versatile emissive range of fluorophores. These fluorophores have been applied to show their efficiency for protein labeling by exploiting the same basic principle. A genetically modified version of class A type β-lactamase has been used as the tag protein (BL-tag). The strength of the aggregation interaction between a fluorophore and a quencher plays a governing role in the elimination step of the quencher from the probes, which ultimately controls the swiftness of the protein labeling strategy. Modulation in the elimination process can be accomplished by the variation in the nature of the fluorophore. This diversity facilitates the study of the competitive binding order among the synthesized probes toward the BL-tag labeling method. An aggregation and elimination-based BL-tag technique has been explored to develop an order of color labeling from the equimolar mixture of the labeling probe in solutions. The qualitative and quantitative determination of ordering within the probes toward labeling studies has been executed through SDS-PAGE and time-dependent fluorescence intensity enhancement measurements, respectively. The desirable multiple-wavelength fluorescence labeling probes for the BL-tag technology have been developed and demonstrate broad applicability of this labeling technology to live cell imaging with coumarin and fluorescein derivatives by using confocal microscopy.

Two cryptand molecules are connected via p-xyloyl, benzene-1,4-dicarbolyol and 9,10-dimethylene anthracene. Each cryptand is further derivatized with fluorophores such that electronic absorption of one fluorophore overlaps emission of the other. This way, three different systems L(1), L(2) and L(3) have been synthesized to get a better view of the effect on the distance in single- and two-step fluorescence resonance energy transfer (FRET) process. These molecules are probed for FRET in the presence of a metal ion as input. L(1) and L(2) exhibit poor performance in single step FRET while in the case of L(3), a large two-step FRET process is operational with Cu(II) or Hg(II) as input.

Attachment of three different fluorophores to an aza-oxa cryptand (L) gives a transition-metal ion induced two-step fluorescence resonance energy transfer system.