Near-Infrared Activation of Sensory Rhodopsin II Mediated by NIR-to-Blue Upconversion Nanoparticles

Direct optical activation of microbial rhodopsins in deep biological tissue suffers from ineffective light delivery because visible light is strongly scattered and absorbed. NIR light has deeper tissue penetration, but NIR-activation requires a transducer that converts NIR light into visible light in proximity to proteins of interest. Lanthanide-doped upconversion nanoparticles (UCNPs) are ideal transducer as they absorb near-infrared (NIR) light and emit visible light. Therefore, UCNP-assisted excitation of microbial rhodopsins with NIR light has been intensively studied by electrophysiology technique. While electrophysiology is a powerful method to test the functional performance of microbial rhodopsins, conformational changes associated with the NIR light illumination in the presence of UCNPs remain poorly understood. Since UCNPs have generally multiple emission peaks at different wavelengths, it is important to reveal if UCNP-generated visible light induces similar structural changes of microbial rhodopsins as conventional visible light illumination does. Here, we synthesize the lanthanide-doped UCNPs that convert NIR light to blue light. Using these NIR-to-blue UCNPs, we monitor the NIR-triggered conformational changes in sensory rhodopsin II from Natronomonas pharaonis (NpSRII), blue light-sensitive microbial rhodospsin, by FTIR spectroscopy. FTIR difference spectrum of NpSRII was recorded under two different excitation conditions: (ⅰ) with conventional blue light, (ⅱ) with UCNP-generated blue light upon NIR excitation. Both spectra display similar spectral features characteristic of the long-lived M photointermediate state during the photocycle of NpSRII. This study demonstrates that NIR-activation of NpSRII mediated by UCNPs takes place in a similar way to direct blue light activation of NpSRII

Influence of the molecular structure of carboxyl-terminated self-assembled monolayer on the electron transfer of cytochrome c adsorbed on an an electrode: In situ observation by surface-enhanced infrared absorption spectroscopy

Jiang X, Ataka K, Heberle J. Influence of the molecular structure of carboxyl-terminated self-assembled monolayer on the electron transfer of cytochrome c adsorbed on an an electrode: In situ observation by surface-enhanced infrared absorption spectroscopy. JOURNAL OF PHYSICAL CHEMISTRY C. 2008;112(3):813-819

Size-Dependent Stability of Water-Solubilized CdTe Quantum Dots and Their Uptake Mechanism by Live HeLa Cells

Water-solubilized quantum dots have led to a promising application in cellular labeling and biological imaging. The physicochemical properties of water-solubilized quantum dots, particularly in a physiological environment, are strongly dependent on their size. In this paper, we systematically studied the stability of mercaptosuccinic acid-coated CdTe quantum dots (MSA-QDs) of about 2.3 and 5.4 nm diameters in various buffers with different pH values and under laser irradiation by fluorescence spectroscopy. It was found that larger MSA-QDs showed better stability. Size-dependent uptake of MSA-QDs by living HeLa cells was further investigated by confocal microscopy. In phosphate buffer solution, the larger MSA-QDs entered the cells mainly by endocytosis, and part of the smaller ones entered the cells by passive penetration. In cell culture medium, their uptake pathways could be changed due to the changes of their surface properties. The cytotoxicity of smaller and larger MSA-QDs was significantly decreased due to the adsorption of some biological components in the cell culture medium on the nanoparticles surface

Breaking of the Phosphodiester Bond: A Key Factor That Induces Hemolysis

In-depth understanding the toxicity of nanomaterials in red blood cells (RBCs) is of great interest, because of the importance of RBCs in transporting oxygen in blood circulation. Although the toxic effects of nanoparticles in RBCs have been revealed, the conclusions from the literature are conflicting, and in particular, the toxic mechanism is still at the infant stage. Herein, we investigated the size-dependent toxicity of well-known CdTe semiconductor quantum dots (QDs) and revealed the exact toxic mechanism at the molecular level by confocal microscopy and Fourier transform infrared (FT-IR) spectroscopy techniques. We found that smaller mercaptosuccinic acid-capped CdTe QDs (MSA-QDs) with the green-emitting color could cause hemagglutination whereas the middle-size yellow-emitting MSA-QDs induced the formation of stomatocytes and echinocytes and the bigger size red-emitting MSA-QDs induced heavy hemolysis and the formation of lots of ghost cells. The FT-IR data proved that all the MSA-QDs were likely to bond to the RBCs membranes and caused the structural changes of lipid and protein in RBCs. But only the red-emitting MSA-QDs caused the breakage of the phosphodiester bond, which might cause the heavy hemolysis. To some extent, this is the first example that reveals the hemolysis mechanism at the molecular level

A Facile One-Pot Synthesis of Copper Sulfide-Decorated Reduced Graphene Oxide Composites for Enhanced Detecting of H₂O₂ in Biological Environments

The high levels of H₂O₂ are closely associated with cancer and progressive neurodegenerative diseases, such as Parkinson’s disease. In this study, we developed a novel CuS nanoparticle-decorated reduced graphene oxide-based electrochemical biosensor for the reliable detection of H₂O₂. The new electrocatalyst, CuS/RGO composites was successfully prepared by heating the mixture of CuCl₂ and Na₂S aqueous solutions in the presence of PVP-protected graphene oxide at 180 °C. A potential application of CuS/RGO composite-modified electrode as a biosensor to monitor H₂O₂ has been investigated. The steady-state current response increases linearly with H₂O₂ concentration from 5 to 1500 μM with a fast response time of less than 2 s. The detection limit (3σ) for determination of H₂O₂ has been estimated to be 0.27 μM, which was lower than certain enzymes and noble metal nanomaterial-based biosensors. In addition, the study of storage time on the amperometric response of the sensor indicates super stability. Due to these remarkable analytical advantages, the as-made sensor was applied to determine the H₂O₂ levels in human serum and urine samples and H₂O₂ released from human cervical cancer cells with satisfactory results. These results demonstrate that this new nanocomposite with the high surface area and electrocatalytic activity is a promising candidate for use as an enhanced electrochemical sensing platform in the design of nonenzymatic biosensors

Cytochrome c superstructure biocomposite nucleated by gold nanoparticle: thermal stability and voltammetric behavior

The thermal stability of cytochrome c (cyt c) after Au-nanoparticle-directed association has been studied by various spectroscopic (electronic absorption, resonance Raman, and circular dichroism) and electrochemical methods. The results show that the thermal stability of the Au-cyt c superstructure biocomposite formed by the electrostatic and hydrophobic interactions among the associated proteins increases significantly. It is mainly caused by strong hydrophobicity of the associated cyt c in Au-cyt c superstructure at high temperature, which results from the compact secondary structure and the packing of hydrophobic side chains around the Trp 59 and heme. In addition, the formation of bis-His configuration of heme is facilitated by the tightly self-associated state of cyt c in the Au-cyt c superstructure. The electrostatic coupling of the opposite charges among shells of the adsorbed proteins due to the formation of the superstructure biocomposite can reduce repulsions among the same charges in protein. These factors are also important for enhancing the stability of the associated cyt c. Furthermore, the voltammetric behavior of Au-cyt c at DNA modified glassy carbon electrode has been investigated for extending the application of Au-cyt c

In situ monitoring of the orientated assembly of strep-tagged membrane proteins on the gold surface by surface enhanced infrared absorption spectroscopy

Jiang X, Zuber A, Heberle J, Ataka K. In situ monitoring of the orientated assembly of strep-tagged membrane proteins on the gold surface by surface enhanced infrared absorption spectroscopy. PHYSICAL CHEMISTRY CHEMICAL PHYSICS. 2008;10(42):6381-6387