An Engineered Biliverdin-Compatible Cyanobacteriochrome Enables a Unique Ultrafast Reversible Photoswitching Pathway

Cyanobacteriochromes (CBCRs) are promising optogenetic tools for their diverse absorption properties with a single compact cofactor-binding domain. We previously uncovered the ultrafast reversible photoswitching dynamics of a red/green photoreceptor AnPixJg2, which binds phycocyanobilin (PCB) that is unavailable in mammalian cells. Biliverdin (BV) is a mammalian cofactor with a similar structure to PCB but exhibits redder absorption. To improve the AnPixJg2 feasibility in mammalian applications, AnPixJg2_BV4 with only four mutations has been engineered to incorporate BV. Herein, we implemented femtosecond transient absorption (fs-TA) and ground state femtosecond stimulated Raman spectroscopy (GS-FSRS) to uncover transient electronic dynamics on molecular time scales and key structural motions responsible for the photoconversion of AnPixJg2_BV4 with PCB (Bpcb) and BV (Bbv) cofactors in comparison with the parent AnPixJg2 (Apcb). Bpcb adopts the same photoconversion scheme as Apcb, while BV4 mutations create a less bulky environment around the cofactor D ring that promotes a faster twist. The engineered Bbv employs a reversible clockwise/counterclockwise photoswitching that requires a two-step twist on ~5 and 35 picosecond (ps) time scales. The primary forward P_fr → P_o transition displays equal amplitude weights between the two processes before reaching a conical intersection. In contrast, the primary reverse P_o → P_fr transition shows a 2:1 weight ratio of the ~35 ps over 5 ps component, implying notable changes to the D-ring-twisting pathway. Moreover, we performed pre-resonance GS-FSRS and quantum calculations to identify the Bbv vibrational marker bands at ~659,797, and 1225 cm⁻¹. These modes reveal a stronger H-bonding network around the BV cofactor A ring with BV4 mutations, corroborating the D-ring-dominant reversible photoswitching pathway in the excited state. Implementation of BV4 mutations in other PCB-binding GAF domains like AnPixJg4, AM1_1870g3, and NpF2164g5 could promote similar efficient reversible photoswitching for more directional bioimaging and optogenetic applications, and inspire other bioengineering advances.     

A phenolic antioxidant from the Pacific oyster (Crassostrea gigas) inhibits oxidation of cultured human hepatocytes mediated by diphenyl-1-pyrenylphosphine

3,5-Dihydroxy-4-methoxybenzyl alcohol (DHMBA), an antioxidant isolated from the Pacific oyster (Crassostrea gigas), was studied in a cell-based fluorometric antioxidant assay using human hepatocyte-derived cells (C3A) and diphenyl-1-pyrenylphosphine (DPPP) as a fluorescent probe. In comparison with two hydrophilic antioxidants, DHMBA showed the stronger inhibition of DPPP-mediated fluorescence than chlorogenic acid and l-ascorbic acid: at a concentration of 320 μM of DPPP, the inhibition was 26.4 ± 2.6%, 11.1 ± 1.2%, and 0 ± 2.0% for DHMBA, chlorogenic acid, and l-ascorbic acid, respectively (mean ± SD, n = 4). Their relative oxygen radical absorbance capacities (ORAC) were dissociated with their cell-based antioxidant activities: 1.47 ± 0.40, 4.57 ± 0.30, and 0.53 ± 0.13 μmol TE/μmol for DHMBA, chlorogenic acid, and l-ascorbic acid, respectively (mean ± SD, n = 4). The amphiphilicity of DHMBA was better than chlorogenic acid and l-ascorbic acid might underlie this dissociation. Since the C3A cells are human hepatoma-derived cells, DHMBA might be useful in the prevention and treatment of liver diseases by involving an oxidation process.     

Optical nonlinearity of Cu:SrTiO3 composite fabricated by negative ion implantation

Optical nonlinearity and dispersion were studied for Cu nanoparticle composite. Negative Cu⁻ ions with 60 keV were applied for implanting into SrTiO₃ at a flux of 10 μA/cm² up to a total fluence of 1 × 10¹⁷ ions/cm². The steady-state optical spectrum of Cu-implanted SrTiO₃ showed a surface plasmon resonance resulting from the formation of nanoparticles. Transient transmission and reflection were measured by the pump–probe method with a femtosecond laser system. Optical nonlinearity originated from Cu:SrTiO₃ nanoparticle composite and from SrTiO₃ matrix overlap, in measured transient spectra. The nonlinear component of Cu:SrTiO₃ composite was extracted from the transient spectra with the difference of time response. Nonlinear dielectric coefficient and dispersion around the surface plasmon resonance were derived from extracted transient spectra. The dispersion was compared with a local electric field factor.     

Magnetic studies of the trinuclear center in laccase and ascorbate oxidase approached by EPR spectroscopy and magnetic susceptibility measurements

The trinuclear centers in Rhus vernicifera laccase and Cucumis sativus ascorbate oxidase have been studied by EPR spectroscopy and magnetic susceptibility measurements over the wide range of 5 K to 300 K. The EPR spectra showed that type II copper receives increasing tetrahedral distortion with raising temperature. Magnetic susceptibilities of laccase showed that both of type I and type II coppers are almost fully paramagnetic since the antiferromagnetic interaction between type III coppers is extremely strong from 5 K to 300 K. On the other hand, the effective magnetic moment of ascorbate oxidase is contributed by ca. 1.7 Cu2+ even below ca. 100 K, since type II Cu is partly in the reduced form. The effective magnetic moment continuously increased with raising temperature because the antiferromagnetic interaction between type III coppers is not as strong as in the case of laccase. The simulation of the SQUID measurement results suggested that the conformational change of the ascorbate oxidase molecule caused the temperature dependence of the antiferromagnetic interaction. The type II Cu EPR signals in laccase and ascorbate oxidase were conspicuously broadened with raising temperature because of the increasing contribution of the triplet state by type III Cu's and/or of the rapid relaxation which finally led to only ca. 30% detection of the type II Cu signals at room temperature. The stepwise binding of azide to the trinuclear center made one of type III Cu's to be EPR detectable. SQUID measurements indicated that only one Cu in the trinuclear center is paramagnetic and other two Cu's are antiferromagnetically coupled for both of the one- and two-azide bound forms. The binding mode of azide to the trinuclear center was discussed based on some models.     

Occurrence of 3-O-methylmannose in the polysaccharide of Leptospira biflexa Urawa

3-O-methylmannose was identified by gas-liquid chromatography-mass spectrometry in the acid hydrolysate of the polysaccharide of Leptospira biflexa Urawa.     

Ionic conductivity of solid lithium ion conductors with the spinel structure: Li2MCl4 (M = Mg,Mn, Fe,Cd)

The ionic conductivity of the polycrystalline samples of the lithium chloride spinel, Li₂MCl₄ (M = Mg, Mn, Fe, Cd) is measured by the help of ac techniques in the temperature range of 25–500°C. The highest conductivity of 0.3 Scm^?1 at 400°C is obtained in Li₂CdCl₄. In addition, the solid solution, Li_2?2δM_1+δCl₄, has been examined. It is found that Li_1.9Cd_1.05Cl₄ has higher conductivity than that of Li₂CdCl₄. The phase transitions of these compounds are also discussed.