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Influence of Arg72 of pharaonis Phoborhodopsin on M-intermediate Decay and Proton Pumping Activity
Ikeura, Yukako,Shimono, Kazumi,Iwamoto, Masayuki,Sudo, Yuki,Kamo, Naoki Korean Society of Photoscience 2002 Journal of Photosciences Vol.9 No.2
X-ray structures of pharaonis phoborhodopsin (ppR) show the different direction of the side chain of Arg72 from that of the corresponding residue (Arg82) of bacteriorhodopsin, BR. For BR, this residue is considered to play an important role in the proton pumping. In order to investigate the role of Arg72 in ppR, we constructed Arg72 mutants of R72A, R72K and R72Q, and measured the photocycle and proton pumping activities. The pH-titration curves on the absorption maximum of the mutants were shifted to alkaline in comparison of that of the wild-type. This may imply the increase of pKa of D75, suggesting the presence of the (probably electric) interaction between D75 and Arg72. Rate constants of the M-decay were 3-7 times faster than that of the wild-type, and the time for the completion of the photocycling was also reduced. Using Sn0$_2$ electrode, the rate of transmembrane proton transport was measured upon illumination. The photo-induced proton pumping activities were estimated after the corrections that are the percentages of the associated form of D75 (which has no pumping activity) and the photocycling rates. R72A and R72Q showed the reduced activity while R72K did not reduce the activity.
Ikeura Atsushi,Kushida Taketoshi,Oe Kenichi,Kotani Yoshihisa,Ando Muneharu,Adachi Takashi,Saito Takanori 대한척추외과학회 2020 Asian Spine Journal Vol.14 No.3
Study Design: Biomechanical study.Purpose: To assess the correlation between the computed tomography (CT) values of the pedicle screw path and screw pull-out strength.Overview of Literature: The correlation between pedicle screw pull-out strength and bone mineral density has been well established. In addition, several reports have demonstrated a correlation between bone mineral density and CT values. However, no previous biomechanical studies investigated the correlation between CT values and pedicle screw pull-out strength.Methods: Sixty fresh-frozen lumbar vertebrae from 6-month-old pigs were used. Before screw insertion, the CT values of the screw path were obtained for each sample. Specimens were then randomly divided into three equal groups. Each group had one of three pedicle screws inserted: 4.0-mm LEGACY (4.0-LEG), 4.5-mm LEGACY (4.5-LEG), or 4.5-mm SOLERA (4.5-SOL) (all from Medtronic Sofamor Danek Inc., Memphis, TN, USA). Each screw had a consistent 30-mm thread length. Axial pull-out testing was performed at a rate of 1.0 mm/min. Correlations between the CT values and pedicle screw pull-out strength were evaluated using Pearson’s correlation coefficient analysis.Results: The correlation coefficients between the CT values of the screw path and pedicle screw pull-out strength for the 4.0-LEG, 4.5-LEG, and 4.5-SOL groups were 0.836 (<i>p</i> <0.001), 0.780 (<i>p</i> <0.001), and 0.873 (<i>p</i> <0.001), respectively. Greater CT values were associated with greater screw pull-out strength.Conclusions: The CT values of the screw path were strongly positively correlated with pedicle screw pull-out strength, regardless of the screw type and diameter, suggesting that the CT values could be clinically useful for predicting pedicle screw pull-out strength.
Shimono, Kazumi,Ikeura, Yukako,Sudo, Yuki,Iwamoto, Masayuki,Kamo, Naoki Korean Society of Photoscience 2002 Journal of Photosciences Vol.9 No.2
pharaonis phoborhodopsin (ppR), a photophobic sensor of haloalkaliphilic bacteria, Natronobacterium phar-aonis, has retinal as a chromophore covalently bound to Lys in G-helix via a protonated Schiff base (PSB), as is the same as bacteriorhodopsin (bR). For ppR, the corresponding counter-ion is Asp residue (Asp75) located in C-helix. Here we investigated the influence of the protonated state of this counter-ion and its location on the chromophore configuration. Under alkaline condition, the chromophore configuration of D75E mutant was analyzed by HPLC. D75E had a much larger content of 13-cis isomer: the ratio of 13-cis to all-trans was 6:4 while the wild-type had this ratio of 1 :9. On the other hand, under acidic condition where Glu was associated, D75E had no 13-cis retinal isomer. Mutants whose Asp75 was replaced by neutral amino acids (D75N and D75Q) did not contain 13-cis retinal. Furthermore, retinal isomer compositions and the change in the visible ab- sorption spectra (indicating the dissociation state of Glu75) were measured under varying pH, and these were almost the same dependencies. These results indicate that an important factor determining the 13-cis isomer content is the presence of negative charge of the counter-ion against PSB, but not the size of this residue. Com- parison between the wild-type and D75E in alkaline solutions indicates the influence of the location of the counter-ion.
Tamaki, Masahiko,Kobayashi, Fumiyuki,Ikeura, Hiromi,Sato, Michio The Korean Society of Plant Pathology 2018 Plant Pathology Journal Vol.34 No.4
To investigate the difference in the disinfectant efficiency of ozone microbubbles ($O_3MB$) and ozone millibubbles ($O_3MMB$), the morphological change of the treated Fusarium oxysporum f. sp. melonis spores was observed with scanning and transmission electron microscopies (SEM and TEM). The disinfectant efficiency of $O_3MB$ on F. oxysporum f. sp. melonis spores was greater than that of $O_3MMB$. On observation with SEM, it was revealed that morphological change of F. oxysporum f. sp. melonis spores was caused by $O_3MB$ and $O_3MMB$, and damage to the spore surfaces by $O_3MB$ occurred sooner than that by $O_3MMB$. On observation with TEM, it was furthermore confirmed that F. oxysporum f. sp. melonis spores treated with $O_3MB$ induced wavy deformation of cell membrane and the intracellular change different from that with $O_3MMB$. Therefore, the greater disinfection efficiency of $O_3MB$ was suggested to be caused due to the function of the MB in addition to the oxidative power of $O_3$.