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Sum frequency generation (SFG) vibrational spectroscopy has been employed to study a variety of interesting biological phenomena occurring at interfaces. This review summarizes recent SFG studies on proteins, lipid monolayers and bilayers, and other biological molecules. Molecular level details revealed by SFG in these studies show that SFG is a powerful technique for characterizing conformation, orientation and ordering of biological molecules at interfaces.

A novel nanofabrication method that combines both "bottom-up" (template-assisted peptide self-assembling) and "top-down" (replica molding) techniques is introduced. A designer peptide, GAV-9 (NH₂-VGGAVVAGV-CONH₂), can epitaxially self-assemble into nanofilaments on the surface of mica, which is further used as the diversified masters for the application of replica molding. With in situ atomic force microscopy monitoring, several typical masters are fabricated by peptide self-assembling on the surface of mica. These masters can be easily molded into hard poly(dimethylsiloxane) surfaces, and then further replica-molded into polyurethane surfaces. The polymeric surfaces with regular 1D and 2D patterns on the nanometer scale are expected to have new applications in nanostructure's fabrication.

Amyloids are stable, β-sheet-rich protein/peptides aggregates with 2–15 nm diameter and few micrometers long. It is originally associated with many human diseases such as Alzheimer's, Parkinson's and prion diseases. Amyloids are resistant to enzyme degradation, temperature changes and wide ranges of pH. Although, amyloids are hard and their stiffness is comparable to steel, a constant recycling of monomer occur inside the amyloid fibrils. It grows in a nucleation dependent polymerization manner by recruiting native soluble protein and by converting them to amyloid. These extraordinary physical properties make amyloids attractive for nanotechnological applications. Some amyloid fibrils have also evolved to perform native biological functions (functional amyloid) of the host organism. Functional amyloids are present in mammals such as amyloids of pMel17 and pituitary hormones, where they help in skin pigmentation and hormone storage, respectively. Here, the progress of utilizing amyloid fibrils for nanobiotechnological applications with particular emphasis on the recent studies that amyloid could be utilized for the formulation of peptide/protein drugs depot and how secretory cells uses amyloid for hormone storage will be reviewed.

In addition to controlling the structure of multi-chromophoric arrays, monitoring the spatial orientation of the chromophores in artificial light harvesting devices is a challenge of growing interest. We report in this article our recent advances in this field. It is expected that a better understanding of the physicochemical properties of rigid cofacial porphyrinic tweezers and an identification of the factors governing them will be crucial for the design and the elaboration of new nano-molecules endowed with original properties. Extended multi-porphyrinic architectures, polypeptides bearing pendant porphyrins have been synthesized as linear devices, and a star-like pentaporphyrin as an arborescent array. The structure and the original conformation of the latter confer to this system an unusual duality in its physicochemical properties.

In this paper we now report our ongoing progress in the preparation of artificial photosynthetic systems through the preparation of light harvesting multi-porphyrins. A tetramer, constituted of a central dipeptide functionalized by two free-base porphyrins and surrounded by one amino-acid bearing a pendant Zn(II) porphyrin on each side, has been chosen. The optical and photophysical properties of this tetramer have been studied by absorption and fluorescence spectroscopy. In addition, the energy transfer phenomenon has been studied and monitored by femtosecond time-resolved fluorescence. Our results indicate that the excited state dynamics redounding in the excitation being localized in the inner free-base porphyrins takes place in the time scale of approximately 1 ps.

We report herein the binding studies of three bis-porphyrinic tweezers bearing nucleosidic linkers and a bis-porphyrinic dipeptide with two bidentate bases of different size, DABCO and 4,4' -bipyridine. In the nucleosidic series, the nucleoside confers, for two dimers out of three, sufficient preorganization to the dimers to enhance the association constants, documenting the fact that rigid dimers are not necessary for preorganization, and opening up new routes to the faster synthesis of flexible tweezers capable of complexing guests with a high association constant. Comparison of these results with those obtained for rigid tweezers shows a better efficiency of the flexible nucleosidic dimers. We thus document the fact that the choice of rigid spacers is not the only way to pre-organize bis-porphyrins, and that some well-chosen nucleosidic linkers offer an interesting option for the synthesis of such devices.

meso-Tris(3,5-di-tert-butylphenyl)porphyrin (P-H) is bonded with L-phenylalanine (H-Phe-OH) directly at the unsubstituted meso-position of the former and the p-position of phenyl group of the latter to afford chiral porphyrin-amino acid conjugate H-Phe(p-P)-OH. The N-(9-fluorenyl)-methyloxycarbonyl compound, Fmoc-Phe(p-P)-OH, was synthesized without any loss of enantiomeric purity (based on chiral HPLC analysis) from commercially available L-tyrosine and was useful for preparation of the peptides in both liquid and solid phases. Other meso-tetraarylporphyrins possessing multi-amino acid moieties are reported, as well as achiral porphyrin-amino acids readily prepared and their dipeptidyl porphyrin dyads.

Five amphiphilic protoporphyrin IX-peptide conjugates bearing the sequences ATWLPPR, AAhexPQRRSARLSA and cERGDPhe conjugated via the propionic side chains, were synthesized and evaluated in vitro using two cell lines: human carcinoma HEp2 and human leukemia HL-60. All conjugates were found to have low dark- and photo-toxicities in both cell lines, and 3 to 10-fold higher accumulation was observed within HL-60 vs. HEp2 cells, depending on the nature of the peptide sequence. The preferential subcellular sites of localization for all conjugates were found to be the lysosomes in HEp2 cells, and the mitochondria in HL-60 cells, suggesting different mechanisms of cellular internalization.

The synthesis and in vitro evaluation of four mesoporphyrin IX-peptide conjugates designed to target EGFR, over-expressed in colorectal and other cancers, are reported. Two peptides with known affinity for EGFR, LARLLT (1) and GYHWYGYTPQNVI (2), were conjugated to mesoporphyrin IX (MPIX, 3) via one or both the propionic side chains, directly (4, 5) or with a triethylene glycol spacer (7, 8). The conjugates were characterized using NMR, MS, CD, SPR, UV-vis and fluorescence spectroscopies. Energy minimization and molecular dynamics suggest different conformations for the conjugates. SPR studies show that conjugate 4, bearing two LARLLT with no PEG spacers, has the greatest affinity for binding to EGFR, followed by conjugate 7with two PEG and two LARLLT sequences. Molecular modeling and docking studies suggest that both conjugates 4 and 7 can bind to monomer and dimer EGFR in open and closed conformations. The cytotoxicity and cellular targeting ability of the conjugates were investigated in human HEp2 cells over-expressing EGFR. All conjugates showed low dark- and photo-toxicities. The cellular uptake was highest for conjugates 4 and 8 and lowest for 7 bearing two LARLLT linked via PEG groups, likely due to decreased hydrophobicity. Among the conjugates investigated, 4 is the most efficient EGFR-targeting agent, and therefore the most promising for the detection of cancers that over-express EGFR.

This short review highlights the author’s group research on modified vitamin B${}_{12}$ derivatives with a peptide backbone as (1) inhibitors of B${}_{12}$-dependent enzymes and as (2) models of cofactor B${}_{12}$-protein complexes.

Porphyrin-peptide conjugates have a breadth of potential applications, including use in photodynamic therapy, boron neutron capture therapy, as fluorescence imaging tags for tracking subcellular localization, as magnetic resonance imaging (MRI) positive-contrast reagents and as biomimetic catalysts. Here, we have explored three general routes to porphyrin-peptide conjugates using the Cu(I)-catalyzed Huisgen-Medal-Sharpless 1,3-dipolar cycloaddition of peptide-containing azides with a terminal alkyne-containing porphyrin, thereby generating porphyrin-peptide conjugates (PPCs) comprised of a cationic porphyrin coupled to short antimicrobial peptides. In addition to characterizing the PPCs using a variety of spectroscopic (UV-vis, ${}^{\mathrm{\text{1}}}$H- and ${}^{\mathrm{\text{13}}}$C-NMR) and mass spectrometric methods, we evaluated their efficacy as photosensitizers for the in vitro photodynamic inactivation of Mycobacterium smegmatis as a model for the pathogen Mycobacterium tuberculosis. Difficulties that needed to be overcome for the efficient synthesis of PPCs were the limited solubility of the quaternized pyridyl porphyrin in common solvents, undesired (de)metallation and transmetallation, and chromatographic purification. Photodynamic inactivation studies of a small library of PPCs against Mycobacterium smegmatis confirmed our hypothesis that the porphyrin-based photosensitizer maintains its ability to efficiently inactivate bacteria when conjugated to a small peptide by upwards of 5–6 log units (99.999$+$%) using white light illumination (400–700 nm, 60 mW/cm${}^{\mathrm{\text{2}}}$, 30 min). Further, hemolysis assays revealed the lack of toxicity of the PPCs against sheep blood at concentrations employed for in vitro photodynamic inactivation. Taken together, the results demonstrated the ability of PPCs to maintain their antimicrobial photodynamic inactivation efficacy when possessing a short cationic peptides for enabling the potential targeting of pathogens in vivo.

The synthesis and biological studies of BODIPY-GPR peptide conjugate (BD-2) are reported. As compared to the parent BODIPY (BD-1), the peptide linked BD-2showed blue shifted absorption and emission with excellent Stokes shift of 201 nm. Molecular docking studies on EGFR protein kinase indicated very efficient binding affinity of BD-2 as compared to the standard drug (Erlotinib). The cell viability experiments of BD-2on normal (HEK293T) and lung cancer (A549) cell lines indicated 85–95% viability. Bioimaging studies showed that, BD-2was able to penetrate the lung cancer cell line.

We demonstrate here a facile and effective strategy to prepare reduced graphene oxide-platinum nanoparticle (RGO-PtNP) nanohybrids by the mediation of graphite-specific peptide (GSP). For the first time, we found that GSP can be used to modify RGO non-covalently in one way, and in another way promote the formation of PtNPs on RGO as a biomolecular bridge. The created RGO-PtNP nanohybrids show enhanced electrocatalytic activity toward H₂O₂ and can be utilized to fabricate non-enzymatic electrochemical H₂O₂ sensor.

Bioinspired synthesis provides a potential green method for creating functional nanomaterials on graphene supports. In this study, we demonstrate the preparation of hedgehog-like cupric oxide nanoclusters (CuONCs) on peptide-modified reduced graphene oxide (RGO-Pep) nanohybrids through a solution-phase synthesis in which the bound peptide molecules (GNNQQNYEE) mediate the non-covalent modification of GO and provide the adsorption of Cu${}^{2+}$ ions and the nucleation sites for the growth of CuONCs. The synthesized RGO-Pep-CuONCs hybrids were further utilized for the modification of a glass carbon electrode to fabricate a non-enzymatic electrochemical sensor for hydrogen peroxide (H₂O₂). It was found that the fabricated H₂O₂ sensor exhibited good performances for sensing H₂O₂ with a detection limit of $0.02\mu$M and two wide linear detection ranges. In addition, this sensor revealed good selectivity and stability. It is expected that the strategies used in this study will be valuable to inspire the creation of various functional biomolecule- and graphene-based hybrid bionanomaterials for the applications in materials science, sensors, biomedical engineering, tissue engineering, nanotechnology, and other fields.