We synthesized Au nanoparticle (AuNP)/ZnO composite particles in presence of an anionic surfactant and evaluated their photocatalytic activity under visible-light irradiation. AuNPs synthesized from ...HAuCl4 in the presence of amylase and the precursor solutions of ZnO were mixed, followed by a hydrothermal process, to synthesize crystal face-controlled AuNP/ZnO composite particles. X-ray diffraction (XRD) patterns and ultraviolet-visible (UV-Vis) spectra confirmed the formation of AuNP/ZnO composite particles. Furthermore, different Au to Zn concentration ratios in the precursor solutions resulted in different amounts of AuNPs in the composite particles. In addition, the average size of the AuNP/ZnO composite particles decreased with increasing Au to Zn concentration ratio. We believe AuNPs act as the nuclei for ZnO particle formation. The photocatalytic activity of the AuNP/ZnO composite particles was evaluated by the photodegradation of methylene blue (MB) under visible-light irradiation. The photodegradation rate of MB was higher with AuNP/ZnO composite particles compared to that with the ZnO particles synthesized in the absence of AuNPs. The AuNP/ZnO composite particles exhibit photocatalytic activity under visible-light irradiation owing to the enhanced charge separation efficiency and the localized surface plasmon resonance effect.
The kidney develops from reciprocal interactions between the metanephric mesenchyme and ureteric bud. The mesenchyme transforms into epithelia and forms complicated nephron structures, whereas the ...ureteric bud extends its pre-existing epithelial ducts. Although the roles are well established for extracellular stimuli, such as Wnt and Notch, it is unclear how the intracellular cytoskeleton regulates these morphogenetic processes. Myh9 and Myh10 encode nonmuscle myosin II heavy chains, and Myh9 mutations in humans are implicated in congenital kidney diseases and focal segmental glomerulosclerosis in adults. Here, we analyzed the roles of Myh9 and Myh10 in the developing kidney. Ureteric bud-specific depletion of Myh9 resulted in no apparent phenotypes, whereas mesenchyme-specific Myh9 deletion caused proximal tubule dilations and renal failure. Mesenchyme-specific Myh9/Myh10 mutant mice died shortly after birth and showed a severe defect in nephron formation. The nascent mutant nephrons failed to form a continuous lumen, which likely resulted from impaired apical constriction of the elongating tubules. In addition, nephron progenitors lacking Myh9/Myh10 or the possible interactor Kif26b were less condensed at midgestation and reduced at birth. Taken together, nonmuscle myosin II regulates the morphogenesis of immature nephrons derived from the metanephric mesenchyme and the maintenance of nephron progenitors. Our data also suggest that Myh9 deletion in mice results in failure to maintain renal tubules but not in glomerulosclerosis.
The candidate tumor suppressor 101F6 protein is a homologue of adrenal chromaffin granule cytochrome b561, which is involved in the electron transfer from cytosolic ascorbate to intravesicular ...monodehydroascorbate radical. Since the tumor suppressor activity of 101F6 was enhanced in the presence of ascorbate, it was suggested that 101F6 might utilize a similar transmembrane electron transfer reaction. Detailed kinetic analyses were conducted on the detergent-solubilized recombinant human 101F6 for its electron transfer reactions with ascorbate and monodehydroascorbate radical by stopped-flow and pulse radiolysis techniques. The reduction of oxidized 101F6 with ascorbate was found to be independent of pH in contrast to those observed for chromaffin granule and Zea mays cytochromes b561 in which both cytochromes exhibited very slow rates at pH 5.0 but faster at pH 6.0 and 7.0. The absence of the inhibition for the electron acceptance from ascorbate upon the treatment with diethyl pyrocarbonate suggested that 101F6 might not utilize a "concerted proton/electron transfer mechanism". The second-order rate constant for the electron donation from the ascorbate-reduced 101F6 to the pulse-generated monodehydroascorbate radical was found to be 5.0 × 10(7) M(-1 )s(-1), about 2-fold faster than that of bovine chromaffin granule cytochrome b561 and about five times faster than that of Zea mays cytochrome b561, suggesting that human 101F6 is very effective for regenerating ascorbate from monodehydroascorbate radical in cells. Present observations suggest that 101F6 employs distinct electron transfer mechanisms on both sides of the membranes from those of other members of cytochrome b561 protein family.
The candidate tumor suppressor 101F6 protein is a homologue of adrenal chromaffin granule cytochrome b 561, which is involved in the electron transfer from cytosolic ascorbate to intravesicular ...monodehydroascorbate radical. Since the tumor suppressor activity of 101F6 was enhanced in the presence of ascorbate, it was suggested that 101F6 might utilize a similar transmembrane electron transfer reaction. Detailed kinetic analyses were conducted on the detergent-solubilized recombinant human 101F6 for its electron transfer reactions with ascorbate and monodehydroascorbate radical by stopped-flow and pulse radiolysis techniques. The reduction of oxidized 101F6 with ascorbate was found to be independent of pH in contrast to those observed for chromaffin granule and Zea mays cytochromes b 561 in which both cytochromes exhibited very slow rates at pH 5.0 but faster at pH 6.0 and 7.0. The absence of the inhibition for the electron acceptance from ascorbate upon the treatment with diethyl pyrocarbonate suggested that 101F6 might not utilize a “concerted proton/electron transfer mechanism”. The second-order rate constant for the electron donation from the ascorbate-reduced 101F6 to the pulse-generated monodehydroascorbate radical was found to be 5.0 × 107 M–1 s–1, about 2-fold faster than that of bovine chromaffin granule cytochrome b 561 and about five times faster than that of Zea mays cytochrome b 561, suggesting that human 101F6 is very effective for regenerating ascorbate from monodehydroascorbate radical in cells. Present observations suggest that 101F6 employs distinct electron transfer mechanisms on both sides of the membranes from those of other members of cytochrome b 561 protein family.
Candidate human tumour suppressor gene product, 101F6 protein, is a highly hydrophobic transmembrane protein and a member of cytochrome b561 family. Purified 101F6 protein expressed in Pichia ...pastoris cells showed visible absorption spectra similar but distinct from those of cytochrome b561. Haem content analysis indicated presence of two haems B per molecule. Midpoint potentials of the purified protein were found as +109 and +26 mV for two haems, slightly lower than those for bovine chromaffin granule or plant Zea mays cytochromes b561. Electron paramagnetic resonance (EPR) spectra in oxidized state at 5 K showed only a highly anisotropic low-spin (HALS) signal at gz = 3.75. However, at 15 and 20 K, another HALS-type signal appeared at gz = 3.65 being overlapped with that of gz = 3.75. The rhombic EPR signal at gz = 3.16 previously seen in other cytochromes b561 was not observed, suggesting distinct haem environments. Absence of the inhibition in the electron transfer from ascorbate by a treatment of 101F6 protein with diethylpyrocarbonate showed a remarkable contrast from those of other cytochromes b561 where the 'concerted H+/e- transfer mechanism' at the cytosolic haem centre was blocked by specific N epsilon -carbethoxylation of haem-coordinating imidazole, suggesting that 101F6 protein might accept electrons via a mechanism distinct from other cytochromes b561.
Candidate human tumour suppressor gene product, 101F6 protein, is a highly hydrophobic transmembrane protein and a member of cytochrome b(561) family. Purified 101F6 protein expressed in Pichia ...pastoris cells showed visible absorption spectra similar but distinct from those of cytochrome b(561). Haem content analysis indicated presence of two haems B per molecule. Midpoint potentials of the purified protein were found as +109 and +26 mV for two haems, slightly lower than those for bovine chromaffin granule or plant Zea mays cytochromes b(561). Electron paramagnetic resonance (EPR) spectra in oxidized state at 5 K showed only a highly anisotropic low-spin (HALS) signal at g(z) = 3.75. However, at 15 and 20 K, another HALS-type signal appeared at g(z) = 3.65 being overlapped with that of g(z) = 3.75. The rhombic EPR signal at g(z) = 3.16 previously seen in other cytochromes b(561) was not observed, suggesting distinct haem environments. Absence of the inhibition in the electron transfer from ascorbate by a treatment of 101F6 protein with diethylpyrocarbonate showed a remarkable contrast from those of other cytochromes b(561) where the 'concerted H(+)/e(-) transfer mechanism' at the cytosolic haem centre was blocked by specific Nε-carbethoxylation of haem-coordinating imidazole, suggesting that 101F6 protein might accept electrons via a mechanism distinct from other cytochromes b(561).
We synthesized Au nanoparticle (AuNP)/ZnO composite particles in presence of an anionic surfactant and evaluated their photocatalytic activity under visible-light irradiation. AuNPs synthesized from ...HAuCl4 in the presence of amylase and the precursor solutions of ZnO were mixed, followed by a hydrothermal process, to synthesize crystal face-controlled AuNP/ZnO composite particles. X-ray diffraction (XRD) patterns and ultraviolet-visible (UV-Vis) spectra confirmed the formation of AuNP/ZnO composite particles. Furthermore, different Au to Zn concentration ratios in the precursor solutions resulted in different amounts of AuNPs in the composite particles. In addition, the average size of the AuNP/ZnO composite particles decreased with increasing Au to Zn concentration ratio. We believe AuNPs act as the nuclei for ZnO particle formation. The photocatalytic activity of the AuNP/ZnO composite particles was evaluated by the photodegradation of methylene blue (MB) under visible-light irradiation. The photodegradation rate of MB was higher with AuNP/ZnO composite particles compared to that with the ZnO particles synthesized in the absence of AuNPs. The AuNP/ZnO composite particles exhibit photocatalytic activity under visible-light irradiation owing to the enhanced charge separation efficiency and the localized surface plasmon resonance effect.
A highly hydrophobic protein with six transmembrane structure that is coded by the candidate tumor suppressor gene 101F6 located in the human chromosome 3p.21.3 and a possible member of the ...cytochrome b 561 protein family was expressed, purified, and characterized in its functional form for the first time. The protein was heterologously expressed in methylotrophic yeast Pichia pastoris as a fusion protein containing a C‐terminal thrombin‐specific sequence and an 8‐His residue tag. Purification was achieved by ion exchange chromatography on DEAE‐Sepharose and affinity chromatography on Ni‐NTA‐Sepharose. SDS‐PAGE analysis revealed a single protein band with an estimated molecular weight of 26 kDa, while Western blot and MALDI‐TOF‐MS analysis confirmed the presence of the cytochrome b561 specific sequence in the protein. The 101F6 protein was found to be reducible by ascorbate efficiently and to have two midpoint potentials at +89.5 and +13.1 mV, slightly lower than the corresponding values of +155 and +62 mV, respectively, of bovine adrenal cytochrome b 561, despite a lower conservation of the putative ascorbate binding site sequence in the 101F6 protein. The “modified motif 1” sequence unique in 101F6 protein may be responsible for other molecular functions, such as protein‐protein interactions, in the endoplasmic membranes.
The candidate tumor suppressor 101F6 protein is a homologue of adrenal chromaffin granule cytochrome b sub(561), which is involved in the electron transfer from cytosolic ascorbate to intravesicular ...monodehydroascorbate radical. Since the tumor suppressor activity of 101F6 was enhanced in the presence of ascorbate, it was suggested that 101F6 might utilize a similar transmembrane electron transfer reaction. Detailed kinetic analyses were conducted on the detergent-solubilized recombinant human 101F6 for its electron transfer reactions with ascorbate and monodehydroascorbate radical by stopped-flow and pulse radiolysis techniques. The reduction of oxidized 101F6 with ascorbate was found to be independent of pH in contrast to those observed for chromaffin granule and Zea mays cytochromes b sub(561) in which both cytochromes exhibited very slow rates at pH 5.0 but faster at pH 6.0 and 7.0. The absence of the inhibition for the electron acceptance from ascorbate upon the treatment with diethyl pyrocarbonate suggested that 101F6 might not utilize a "concerted proton/electron transfer mechanism". The second-order rate constant for the electron donation from the ascorbate-reduced 101F6 to the pulse-generated monodehydroascorbate radical was found to be 5.0 10 super(7) M super(-1 )s super(-1), about 2-fold faster than that of bovine chromaffin granule cytochrome b sub(561) and about five times faster than that of Zea mays cytochrome b sub(561), suggesting that human 101F6 is very effective for regenerating ascorbate from monodehydroascorbate radical in cells. Present observations suggest that 101F6 employs distinct electron transfer mechanisms on both sides of the membranes from those of other members of cytochrome b sub(561) protein family.
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