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101.
Rational Design of Domain‐Swapping‐Based c‐Type Cytochrome Heterodimers by Using Chimeric Proteins 下载免费PDF全文
Mohan Zhang Tsukasa Nakanishi Dr. Masaru Yamanaka Dr. Satoshi Nagao Dr. Sachiko Yanagisawa Dr. Yasuhito Shomura Dr. Naoki Shibata Prof. Dr. Takashi Ogura Prof. Dr. Yoshiki Higuchi Prof. Dr. Shun Hirota 《Chembiochem : a European journal of chemical biology》2017,18(17):1712-1715
The design of protein oligomers with multiple active sites has been gaining interest, owing to their potential use for biomaterials, which has encouraged researchers to develop a new design method. Three‐dimensional domain swapping is the unique phenomenon in which protein molecules exchange the same structural region between each other. Herein, to construct oligomeric heme proteins with different active sites by utilizing domain swapping, two c‐type cytochrome‐based chimeric proteins have been constructed and the domains swapped. According to X‐ray crystallographic analysis, the two chimeric proteins formed a domain‐swapped dimer with two His/Met coordinated hemes. By mutating the heme coordination structure of one of the two chimeric proteins, a domainswapped heterodimer with His/Met and His/H2O coordinated hemes was formed. Binding of an oxygen molecule to the His/H2O site of the heterodimer was confirmed by resonance Raman spectroscopy, in which the Fe?O2 stretching band was observed at 580 cm?1 for the reduced/oxygenated heterodimer (at 554 cm?1 under an 18O2 atmosphere). These results show that domain swapping is a useful method to design multiheme proteins. 相似文献
102.
Sachiko Hirobe Keisuke Imaeda Masashi Tachibana Naoki Okada 《International journal of molecular sciences》2022,23(7)
To improve the efficacy and safety of chimeric antigen receptor (CAR)-expressing T cell therapeutics through enhanced CAR design, we analysed CAR structural factors that affect CAR-T cell function. We studied the effects of disulphide bonding at cysteine residues and glycosylation in the HD on CAR-T function. We used first-generation CAR[V/28/28/3z] and CAR[V/8a/8a/3z], consisting of a mouse vascular endothelial growth factor receptor 2 (VEGFR2)-specific single-chain variable fragment tandemly linked to CD28- or CD8α-derived HD, transmembrane domain (TMD) and a CD3ζ-derived signal transduction domain (STD). We constructed structural variants by substituting cysteine with alanine and asparagine (putative N-linked glycosylation sites) with aspartate. CAR[V/28/28/3z] and CAR[V/8a/8a/3z] formed homodimers, the former through a single HD cysteine residue and the latter through the more TMD-proximal of the two cysteine residues. The absence of disulphide bonds did not affect membrane CAR expression but reduced antigen-specific cytokine production and cytotoxic activity. CAR[V/28/28/3z] and CAR[V/8a/8a/3z] harboured one N-linked glycosylation site, and CAR[V/8a/8a/3z] underwent considerable O-linked glycosylation at an unknown site. Thus, N-linked glycosylation of CAR[V/28/28/3z] promotes stable membrane CAR expression, while having no effect on the expression or CAR-T cell activity of CAR[V/8a/8a/3z]. Our findings demonstrate that post-translational modifications of the CAR HD influence CAR-T cell activity, establishing a basis for future CAR design. 相似文献
103.
Sachiko Murase Sarah E. Robertson Crystal L. Lantz Ji Liu Daniel E. Winkowski Elizabeth M. Quinlan 《International journal of molecular sciences》2022,23(5)
The deletion of matrix metalloproteinase MMP9 is combined here with chronic monocular deprivation (cMD) to identify the contributions of this proteinase to plasticity in the visual system. Calcium imaging of supragranular neurons of the binocular region of primary visual cortex (V1b) of wild-type mice revealed that cMD initiated at eye opening significantly decreased the strength of deprived-eye visual responses to all stimulus contrasts and spatial frequencies. cMD did not change the selectivity of V1b neurons for the spatial frequency, but orientation selectivity was higher in low spatial frequency-tuned neurons, and orientation and direction selectivity were lower in high spatial frequency-tuned neurons. Constitutive deletion of MMP9 did not impact the stimulus selectivity of V1b neurons, including ocular preference and tuning for spatial frequency, orientation, and direction. However, MMP9−/− mice were completely insensitive to plasticity engaged by cMD, such that the strength of the visual responses evoked by deprived-eye stimulation was maintained across all stimulus contrasts, orientations, directions, and spatial frequencies. Other forms of experience-dependent plasticity, including stimulus selective response potentiation, were normal in MMP9−/− mice. Thus, MMP9 activity is dispensable for many forms of activity-dependent plasticity in the mouse visual system, but is obligatory for the plasticity engaged by cMD. 相似文献
104.
Influence of pH and flow on the polarisation behaviour of pure magnesium in borate buffer solutions 总被引:1,自引:0,他引:1
Sachiko Hiromoto Akiko Yamamoto Norio Maruyama Hidetoshi Somekawa Toshiji Mukai 《Corrosion Science》2008,50(12):3561-3568
The influence of pH and flow on the polarisation and impedance behaviour of pure Mg was investigated in 0.6 wt% NaCl borate buffer solutions at pH 6.7, 7.6, and 9.3 using a rotating electrode. The existence of flow prevented the accumulation of the corrosion product and promoted uniform corrosion, leading to an increase of the anodic current density and a decrease of the impedance regardless of the pH of the bulk solution. The influence of the pH did not obviously appear under the static condition, whereas, under the dynamic condition, the anodic current density at pH 6.7 and 7.6 was higher than that at pH 9.3. This result suggests that pH near the surface can be buffered at the pH of bulk solution by borate buffer with the acceleration of ion diffusion. 相似文献
105.
S. M. Smith and D. R. Tindell (1997) reported that prior study of words that are orthographically similar to the solutions of test word fragments (e.g., studying ANALOGY and completing the fragment A L_ _GY, whose solution is ALLERGY) reduced the fragment completion rate relative to a baseline condition in which unrelated words were studied. They called this effect the memory-block effect. In the present experiment, the authors replicated this effect using a larger set of materials than that used by S. M. Smith and D. R. Tindell. The authors also found that dividing attention at study eliminated the memory-block effect. This pattern mimicked the effect of dividing attention on recognition memory but differed from the effect on repetition priming effects. The authors suggest that the memory-block effect is driven by a mechanism different from that responsible for producing repetition priming effects in an implicit fragment completion test. (PsycINFO Database Record (c) 2010 APA, all rights reserved) 相似文献
106.
Manami Semba Shinji Takamatsu Sachiko Komazawa-Sakon Eiji Miyoshi Chiharu Nishiyama Hiroyasu Nakano Kenta Moriwaki 《International journal of molecular sciences》2022,23(13)
Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a cytotoxic cytokine that induces cancer cell death by binding to TRAIL receptors. Because of its selective cytotoxicity toward cancer cells, TRAIL therapeutics, such as recombinant TRAIL and agonistic antibodies targeting TRAIL receptors, have garnered attention as promising cancer treatment agents. However, many cancer cells acquire resistance to TRAIL-induced cell death. To overcome this issue, we searched for agents to sensitize cancer cells to TRAIL-induced cell death by screening a small-molecule chemical library consisting of diverse compounds. We identified a cardiac glycoside, proscillaridin A, as the most effective TRAIL sensitizer in colon cancer cells. Proscillaridin A synergistically enhanced TRAIL-induced cell death in TRAIL-sensitive and -resistant colon cancer cells. Additionally, proscillaridin A enhanced cell death in cells treated with TRAIL and TRAIL sensitizer, the second mitochondria-derived activator of caspase mimetic. Proscillaridin A upregulated TRAIL receptor expression, while downregulating the levels of the anti-cell death molecules, cellular FADD-like IL-1β converting enzyme-like inhibitor protein and Mcl1, in a cell type-dependent manner. Furthermore, proscillaridin A enhanced TRAIL-induced cell death partly via O-glycosylation. Taken together, our findings suggest that proscillaridin A is a promising agent that enhances the anti-cancer efficacy of TRAIL therapeutics. 相似文献
107.
Epigallocatechin gallate (EGCG) must be released in a controlled manner in order to maximize its long-term potential as an antioxidant. In this study, lipase-catalyzed polymerization of EGCG with divinyl adipate and sugar alcohols was conducted to form polyEGCG, which enables long-term controlled release of EGCG via hydrolysis of ester linkages. The polymer yields were influenced by monomer ratios, volume of solvent, and lipase concentration. EGCG was gradually released from polyEGCG under a physiological condition as ester groups of polyEGCG degraded over a period of more than two months, and the releasing rate was influenced by the type of the sugar alcohols and EGCG ratios. EGCG released from polyEGCG maintained its antioxidant activity and the cell viability of polyEGCG eluent with polymer concentration higher than 0.05 mg/mL was as high as 100%. These results indicate that polyEGCG has potential application as a highly efficient and long-term antioxidative material with low cytotoxicity. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019 , 136, 47693. 相似文献
108.
Sayaka Ishihara Makoto Nakauma Takahiro Funami Sachiko Odake Katsuyoshi Nishinari 《Food Hydrocolloids》2011
Swallowing profiles of food polysaccharide gels were investigated in relation to bolus rheology. Polysaccharide gel from either gellan gum or a mixture of gellan gum and psyllium seed gum was used as a model food. Acoustic analysis and sensory evaluation were carried out to investigate the swallowing profiles using the same human subjects. Model bolus was prepared through instrumental mastication using a mechanical simulator to mimic the action of the human jaw in the presence or absence of artificial saliva and was subjected to dynamic viscoelasticity measurements to investigate the rheological properties. Bolus from the binary gel was shorter in time required to transfer through the pharyngeal phase due to mass flow and was scored higher in sensory perceived cohesiveness (bolus forming) than that from gellan gum gel. Model bolus from the binary gel showed a rheologically weak gel (or structured fluid) behavior and was higher in structural homogeneity than that from gellan gum gel. Also, dynamic viscoelasticity parameters of the binary gel were less dependent on the addition level of saliva. Results indicate that the viscoelasticity balance is a key for texture design of dysphagia foods in relation to the saliva miscibility. 相似文献
109.
Time-evolutions of two-level and multi-level systems in phase relaxation processes are investigated in a systematic way. A general form of the decoherence function which determines phase relaxation of a quantum state is obtained by making use of a microscopic interaction Hamiltonian between the relevant system and reservoir. Furthermore the loss of the phase coherence is described in terms of the Pegg–Barnett phase operator. The decoherence function is explicitly calculated for the stochastic dephasing, the boson detector model and the Coleman–Hepp model. 相似文献
110.
Akira Nakajima Tomoki Kobayashi Toshihiro Isobe Sachiko Matsushita 《Materials Letters》2011,65(19-20):3051-3054
Porous spherical CeO2 particles were prepared by impregnation of a cerium precursor solution into organic monolith sphere particles, with subsequent firing at 500 °C in air. The single-phase CeO2 powder had specific surface area of greater than 140 m2/g. Photodeposition with UV illumination loaded Au onto the CeO2 particle surface, which changed from yellowish to purple because of localized surface plasmon resonance (LSPR). The Au-loading increased photocatalytic decomposition activity of the CeO2 powder for gaseous 2-propanol (IPA) under visible light. Thermal desorption of IPA, which was adsorbed to all porous spheres, provided flux to the photocatalytic reaction field of the sphere outer surface. 相似文献