Peptide Nanomaterials Designed from Natural Supramolecular Systems

Natural supramolecular assemblies exhibit unique structural and functional properties that have been optimized over the course of evolution. Inspired by these natural systems, various bio-nanomaterials have been developed using peptides, proteins, and nucleic acids as components. Peptides are attractive building blocks because they enable the important domains of natural protein assemblies to be isolated and optimized while retaining the original structures and functions. Furthermore, the peptide subunits can be conjugated with exogenous molecules such as peptides, proteins, nucleic acids, and metal nanoparticles to generate advanced functions. In this personal account, we summarize recent progress in the construction of peptide-based nanomaterial designed from natural supramolecular systems, including (1) artificial viral capsids, (2) self-assembled nanofibers, and (3) protein-binding motifs. The peptides inspired by nature should provide new design principles for bio-nanomaterials

XAFS analyses of molten metal fluorides

X-ray absorption fine structure studies of molten metal fluorides containing the materials related to nuclear engineering are intensively summarized. By using XAFS spectra data of divalent and trivalent cation metal fluorides in molten state which have been collected by authors’ group for a few years, local structure have been extracted and discussed systematically in conjunction with other spectroscopic studies and numerical calculations. In molten divalent fluorides, tetrahedral coordination of fluorides around a cation is predominant. In the case of pure molten trivalent fluorides, structure with more than 6-coordination has been suggested in some cases, but octahedral coordination structure is much stabilized at heavier rare earth metal fluorides. By mixing with alkali metal fluorides, it is a general trend that inter-ionic distances keep constant, but coordination number of fluorides decreases. In experimental chapter, all the details of sample preparation, furnace installation, X-ray optics setups and data analyses procedures are explained. Finally, future expectations of XAFS technique are also suggested

Construction of Artificial Viral Capsids Encapsulating Short DNAs via Disulfide Bonds and Controlled Release of DNAs by Reduction

To construct an artificial viral capsid encapsulated short single-stranded DNA, a β-annulus peptide conjugated with ssDNA through a disulfide bond at the N-terminus (DNA-SS-β-Annulus) was synthesized. The DNA-SS-β-Annulus conjugate self-assembled into spherical structures ranging in the size of 36–60 nm. ssDNA was released from the capsids via the reduction of disulfide bonds

Squalene Quantification Using Octadecylbenzene as the Internal Standard

AbstractSqualene was determined by HPLC using octadecylbenzene (ODB) as an internal standard. Squalene and ODB were monitored at 210nm. The retention times of squalene and ODB were 7.59 and 8.54min, respectively. Squalene was determined from the peak area ratios of squalene/ODB detected at 210nm. After treatment with 0.5M KOH containing ethanol at 90°C for 1h, squalene in the saponified lipid fraction was extracted using n-hexane. No interfering peak was observed. Linearity of this method was observed in the range 80–900 ng. ODB is useful as an internal standard for squalene determinations

Development of the analog ASIC for multi-channel readout X-ray CCD camera

We report on the performance of an analog application-specific integrated circuit (ASIC) developed aiming for the front-end electronics of the X-ray CCDcamera system onboard the next X-ray astronomical satellite, ASTRO-H. It has four identical channels that simultaneously process the CCD signals. Distinctive capability of analog-to-digital conversion enables us to construct a CCD camera body that outputs only digital signals. As the result of the front-end electronics test, it works properly with low input noise of =<30 uV at the pixel rate below 100 kHz. The power consumption is sufficiently low of about 150 mW/chip. The input signal range of 720 mV covers the effective energy range of the typical X-ray photon counting CCD (up to 20 keV). The integrated non-linearity is 0.2% that is similar as those of the conventional CCDs in orbit. We also performed a radiation tolerance test against the total ionizing dose (TID) effect and the single event effect. The irradiation test using 60Co and proton beam showed that the ASIC has the sufficient tolerance against TID up to 200 krad, which absolutely exceeds the expected amount of dose during the period of operating in a low-inclination low-earth orbit. The irradiation of Fe ions with the fluence of 5.2x10^8 Ion/cm2 resulted in no single event latchup (SEL), although there were some possible single event upsets. The threshold against SEL is higher than 1.68 MeV cm^2/mg, which is sufficiently high enough that the SEL event should not be one of major causes of instrument downtime in orbit.Comment: 16 pages, 6 figure

Horseradish Peroxidase-Decorated Artificial Viral Capsid Constructed from β-Annulus Peptide via Interaction between His-Tag and Ni-NTA

Artificial construction of spherical protein assemblies has attracted considerable attention due to its potential use in nanocontainers, nanocarriers, and nanoreactors. In this work, we demonstrate a novel strategy to construct peptide nanocapsules (artificial viral capsids) decorated with enzymes via interactions between His-tag and Ni-NTA. A β-annulus peptide derived from the tomato bushy stunt virus was modified with Ni-NTA at the C-terminus, which is directed toward the exterior surface of the artificial viral capsid. The β-annulus peptide bearing Ni-NTA at the C-terminus self-assembled into capsids of about 50 nm in diameter. The Ni-NTA-displayed capsids were complexed with recombinant horseradish peroxidase (HRP) with a C-terminal His-tag which was expressed in Escherichia coli. The β-annulus peptide-HRP complex formed spherical assemblies whose sizes were 30–90 nm, with the ζ-potential revealing that the HRP was decorated on the outer surface of the capsid

Dramatic morphological changes in liposomes induced by peptide nanofibers reversibly polymerized and depolymerized by the photoisomerization of spiropyran

Cytoskeletons such as microtubules and actin filaments are natural protein assemblies, which dynamically control cellular morphology by reversible polymerization/depolymerization. Recently, the control of polymerization/depolymerization of fibrous protein/peptide assemblies by external stimuli has attracted significant attention. However, as far as we know, the creation of an “artificial cytoskeleton” that reversibly controls the polymerization/depolymerization of peptide nanofiber in giant unilamellar vesicles (GUVs) has not been reported. Here, we developed peptide nanofiber self-assembled from spiropyran (SP)-modified β-sheet-forming peptides, which can be reversibly polymerized/depolymerized by light. The reversible photoisomerization of the SP-modified peptide (FKFECSPKFE) to the merocyanine-peptide (FKFECMCKFE) by ultraviolet (UV) and visible light irradiation was confirmed by UV–visible spectroscopy. Confocal laser scanning microscopy with thioflavin T staining and transmission electron microscopy of the peptides showed that the SP-peptide formed β-sheet nanofibers, whereas the photoisomerization to the merocyanine-peptide almost completely dissociated the nanofibers. The merocyanine peptide was encapsulated in spherical GUVs comprising of phospholipids as artificial cell models. Interestingly, the morphology of GUV encapsulating the merocyanine-peptide dramatically changed into worm-like vesicles by the photoisomerization to the SP-modified peptide, and then reversibly changed into spherical GUV by the photoisomerization to the MC-modified peptide. These dynamic morphological changes in GUVs by light can be applied as components of a molecular robot with artificially controlled cellular functions

An Analysis of Consistency of Scientific Concept : Based on the Actual Conditions of Undergraduate School Students

The purpose of this study is to provide some implications for curriculum and instruction of buoyancy in secondary school science. The paper-test items about buoyancy in liquids based on the research of Zeineddin & Abd-El-Khalick (2010) were administered to 159 university students. The results of 91% students who had learned physics at high school showed the difficulty of description/application of the theory of buoyancy at different contexts. Although they described the theory of buoyancy, some students can’t perform scientific reasoning well at the other context. These difficulties are caused by memorizing the formula or rules without the understanding of conception of buoyancy in liquids.本研究はJSPS科研費25242015の助成を受けたものである