Browsing by Author "Chen, Shih-Hsun"

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Combined Experimental and Theoretical Study of Long-Range Interactions Modulating Dimerization and Activity of Yeast Geranylgeranyl Diphosphate Synthase
(2009-03-25)
Lo, Chia-Hsiang
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Chang, Ying-Hsuan
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Wright, Jon D.
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Chen, Shih-Hsun
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Kan, Daphne
We present here how two amino acid residues in the first helix distal from the main dimer interface modulate the dimerization and activity of a geranylgeranyl diphosphate synthase (GGPPs). The enzyme catalyzes condensation of farnesyl diphosphate and isopentenyl diphosphate to generate a C(20) product as a precursor for chlorophylls, carotenoids, and geranylgeranylated proteins. The 3D structure of GGPPs from Saccharomyces cerevisiae reveals an unique positioning of the N-terminal helix A, which protrudes into the other subunit and stabilizes dimerization, although it is far from the main dimer interface. Through a series of mutants that were characterized by analytic ultracentrifugation (AUC), the replacement of L8 and 19 at this helix with Gly was found sufficient to disrupt the dimer into a monomer, leading to at least 10(3)-fold reduction in activity. Molecular dynamics simulations and free energy decomposition analyses revealed the possible effects of the mutations on the protein structures and several critical interactions for maintaining dimerization. Further site-directed mutagenesis and AUC studies elucidated the molecular mechanism for modulating dimerization and activity by long-range interactions.
Conjugation of Benzylvanillin and Benzimidazole Structure Improves DNA Binding with Enhanced Antileukemic Properties
(2013-11-15)
Al-Mudaris, Zena A.
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Majid, Aman S. A.
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Ji, Dan
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Al-Mudarris, Ban A.
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Chen, Shih-Hsun
Benzyl-o-vanillin and benzimidazole nucleus serve as important pharmacophore in drug discovery. The benzyl vanillin (2-(benzyloxy)-3-methoxybenzaldehyde) compound shows anti-proliferative activity in HL60 leukemia cancer cells and can effect cell cycle progression at G2/M phase. Its apoptosis activity was due to disruption of mitochondrial functioning. In this study, we have studied a series of compounds consisting of benzyl vanillin and benzimidazole structures. We hypothesize that by fusing these two structures we can produce compounds that have better anticancer activity with improved specificity particularly towards the leukemia cell line. Here we explored the anticancer activity of three compounds namely 2-(2-benzyloxy-3-methoxyphenyl)-1H-benzimidazole, 2MP, N-1-(2-benzyloxy-3-methoxybenzyl)-2-(2-benzyloxy-3-methoxyphenyl)-1H-benzimidazole, 2XP, and (R) and (S)-1-(2-benzyloxy-3-methoxyphenyl)-2, 2, 2-trichloroethyl benzenesulfonate, 3BS and compared their activity to 2-benzyloxy-3-methoxybenzaldehyde, (Bn1), the parent compound. 2XP and 3BS induces cell death of U937 leukemic cell line through DNA fragmentation that lead to the intrinsic caspase 9 activation. DNA binding study primarily by the equilibrium binding titration assay followed by the Viscosity study reveal the DNA binding through groove region with intrinsic binding constant 7.39 mu M/bp and 6.86 mu M/bp for 3BS and 2XP respectively. 2XP and 3BS showed strong DNA binding activity by the UV titration method with the computational drug modeling showed that both 2XP and 3BS failed to form any electrostatic linkages except via hydrophobic interaction through the minor groove region of the nucleic acid. The benzylvanillin alone (Bn1) has weak anticancer activity even after it was combined with the benzimidazole (2MP), but after addition of another benzylvanillin structure (2XP), stronger activity was observed. Also, the combination of benzylvanillin with benzenesulfonate (3BS) significantly improved the anticancer activity of Bn1. The present study provides a new insight of benzyl vanillin derivatives as potential anti-leukemic agent.
Fabrication and characterization of eutectic bismuth-tin (Bi-Sn) nanowires
(2009-05-31)
Chen, Shih-Hsun
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Chen, Chien-Chon
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Luo, Z. P.
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Chao, Chuen-Guang
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材料科學與工程學系
Eutectic Bi-43Sn (in weight percent) nanowires with diameters of 20 nm, 70 nm and 220 nm respectively, were fabricated by a hydraulic pressure injection process using anodic aluminum oxide (AAO) as templates. Novel eutectic microstructure was found within the fabricated nanowires. which are composed of alternating Bi and Sn segments along their wire axes. Within the segments, the electron diffraction analysis revealed single crystalline structures of Bi and Sn elements respectively. Parameters that control the nanowire fabrication process were discussed. It was found out that as the wire diameter reduced, longer Bi and Sn segments formed. (C) 2009 Elsevier B.V. All rights reserved.
Fabrication of bismuth oxide-tin oxide nanowires by direct thermal oxidation of Bi-Sn eutectic nanowires
(2010-11-30)
Chen, Shih-Hsun
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Wang, Chiu-Yen
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Liu, Tzeng-Feng
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Chao, Chuen-Guang
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材料科學與工程學系
Bismuth oxide-tin oxide (BiO(x)-SnO(x)) heterostructure nanowires with a diameter of 70 nm were fabricated by directly annealing Bi-Sn eutectic nanowires synthesized by the vacuum hydraulic pressure injection process. After removal of AAO (Anodic Aluminum Oxide) template with an etching solution, a spontaneous oxide was formed on nanowires to enclose the Bi-Sn eutectic alloys While these nanowires went through the annealing process with the proper heating rate of 50 degrees C/min, the well-annealed oxide nanowires remain solid, straight and segmental The results of cathodoluminescence (CL) spectrum and photoresponse proved that the products consisted of bismuth oxide and tin oxide This fabrication methodology provides a simple way to produce one-dimensional oxide nanomatenals. (C) 2010 Elsevier B.V. All rights reserved
Fabrication of pure aluminum nanowires by using injection molding process in ambient air
(2015-06-01)
Chen, Shih-Hsun
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Wang, Chiu-Yen
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Chen, Yung-Chang
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Hun, Chien-Wan
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Chen, Shih-Fan
A simple method to prepare aluminum nanowires was achieved by using the injection molding process in the air. The Al nanowires were made with the average diameter of 60 nm and length of several micrometers. The difference between mold dimension and nanowires was caused by removing the outer oxide shell of Al nanowires. XRD, EDS and SAED results showed that the Al crystalline nanowires were fabricated exactly. The proposed concept successfully established a simple and low-cost method to have high-temperature and soft nano-materials under the non-vacuum surroundings. (C) 2015 Elsevier B.V. All rights reserved.
Highly Sensitive Arrayed Indium-Antimony Nanowires for Infrared Detection
(2015-01-01)
Chen, Po-Chun
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Chen, Chien-Chon
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Chen, Shih-Hsun
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Chou, Chung-Yi
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Hsieh, Sheng-Jen
The objective of this study is to achieve a high sensitive infrared detector by fabricating highly ordered array of indium-antimony (In-Sb) nanowires which is a semiconductor material. The approach is to investigate an infrared detector with arrayed nanowires which can transport signals in one dimension to obtain high efficiency and sensitivity compared with In-Sb by using traditional thin film fabrications. This research expects to provide an infrared detector by fabricating III-V alloy nanowires to highly improve the resolution of infrared signal. To develop scaled-up functional devices, highly ordered nanowire arrays are essential building blocks. Many candidate materials (metals, alloys, oxides and semiconductors) have been studied for various potential applications in nanotechnology and have shown some promising results. The solid metallic nanowires have been exploited for a wide range of applications to take the advantages of their large length/diameter aspect ratio. Further development to synthesize nanowires efficiently at lower cost is the direction for manufacturing next generation nanodevices. In this study, various diameters of ordering nanowires, from 10 nm to 500 nm, were fabricated and evaluated the performance of the sensitivity of infrared detection. Moreover, a 1 inch plate, which can be regarded as a device, with nanowires array was fabricated by designing a new type of processing chamber.
Moiety-Linkage Map Reveals Selective Nonbisphosphonate Inhibitors of Human Geranylgeranyl Diphosphate Synthase
(2013-09-01)
Chen, Shih-Hsun
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Lin, Sheng-Wei
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Lin, Shen-Rong
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Liang, Po-Huang
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Yang, Jinn-Moon
Bisphosphonates are potent inhibitors of farnesyl pyrophosphate synthase (FPPS) and geranylgeranyl diphosphate synthase (GGPPS). Current bisphosphonate drugs (e.g., Fosamax and Zometa) are highly efficacious in the treatment of bone diseases such as osteoporosis, Paget's disease, and tumor-induced osteolysis, but they are often less potent in blood and soft-tissue due to their phosphate moieties. The discovery of nonbisphosphonate inhibitors of FPPS and/or GGPPS for the treatment of bone diseases and cancers is, therefore, a current goal. Here, we propose a moiety-linkage-based method, combining a site-moiety map with chemical structure rules. (CSRs), to discover nonbisphosphonate inhibitors from thousands of commercially available compounds and known crystal structures. Our moiety-linkage map reveals the binding mechanisms and inhibitory efficacies of 51 human GGPPS (hGGPPS) inhibitors. To the best of our knowledge, we are the first team to discover two novel selective nonbisphosphonate inhibitors, which bind to the inhibitory site of hGGPPS, using CSRs and site-moiety maps. These two compounds can be considered as a novel lead for the potent inhibitors of hGGPPS for the treatment of cancers and mevalonate-pathway diseases. Moreover, based on our moiety-linkage map, we identified two key residues of hGGPPS, K202, and K212, which play an important role for the inhibitory effect of zoledronate (IC50 = 3.4 mu M and 2.4 mu M, respectively). This result suggests that our method can discover specific hGGPPS inhibitors across multiple prenyltransferases. These results show that the compounds that highly fit our moiety-linkage map often inhibit hGGPPS activity and induce tumor cell apoptosis. We believe that our method is useful for discovering potential inhibitors and binding mechanisms for pharmaceutical targets.
New Flexible Silicone-Based EEG Dry Sensor Material Compositions Exhibiting Improvements in Lifespan, Conductivity, and Reliability
(2016-11-01)
Yu, Yi-Hsin
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Chen, Shih-Hsun
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Chang, Che-Lun
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Lin, Chin-Teng
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Hairston, W. David
This study investigates alternative material compositions for flexible silicone-based dry electroencephalography (EEG) electrodes to improve the performance lifespan while maintaining high-fidelity transmission of EEG signals. Electrode materials were fabricated with varying concentrations of silver-coated silica and silver flakes to evaluate their electrical, mechanical, and EEG transmission performance. Scanning electron microscope (SEM) analysis of the initial electrode development identified some weak points in the sensors' construction, including particle pull-out and ablation of the silver coating on the silica filler. The newly-developed sensor materials achieved significant improvement in EEG measurements while maintaining the advantages of previous silicone-based electrodes, including flexibility and non-toxicity. The experimental results indicated that the proposed electrodes maintained suitable performance even after exposure to temperature fluctuations, 85% relative humidity, and enhanced corrosion conditions demonstrating improvements in the environmental stability. Fabricated flat (forehead) and acicular (hairy sites) electrodes composed of the optimum identified formulation exhibited low impedance and reliable EEG measurement; some initial human experiments demonstrate the feasibility of using these silicone-based electrodes for typical lab data collection applications.
Roles of Amino Acids in the Escherichia coli Octaprenyl Diphosphate Synthase Active Site Probed by Structure-Guided Site-Directed Mutagenesis
(2012-04-24)
Chang, Keng-Ming
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Chen, Shih-Hsun
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Kuo, Chih-Jung
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Chang, Chi-Kang
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Guo, Rey-Ting
Octaprenyl diphosphate synthase (OPPS) catalyzes consecutive condensation reactions of farnesyl diphosphate (FPP) with five molecules of isopentenyl diphosphates (IPP) to generate C-40 octaprenyl diphosphate, which constitutes the side chain of ubiquinone or menaquinone. To understand the roles of active site amino acids in substrate binding and catalysis, we conducted site-directed mutagenesis studies with Escherichia coli OPPS. In conclusion, D85 is the most important residue in the first DDXXD motif for both FPP and IPP binding through an H-bond network involving R93 and R94, respectively, whereas R94, K45, R48, and H77 are responsible for IPP binding by providing H-bonds and ionic interactions. K170 and T171 may stabilize the farnesyl carbocation intermediate to facilitate the reaction, whereas R93 and K225 may stabilize the catalytic base (MgPPi) for H-R proton abstraction after IPP condensation. K225 and K235 in a flexible loop may interact with FPP when the enzyme becomes a closed conformation, which is therefore crucial for catalysis. Q208 is near the hydrophobic part of IPP and is important for IPP binding and catalysis.
Targeting the XIAP/caspase-7 complex selectively kills caspase-3-deficient malignancies
(2013-09-01)
Lin, Yuan-Feng
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Lai, Tsung-Ching
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Chang, Chih-Kang
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Chen, Chi-Long
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Huang, Ming-Shyan
Caspase-3 downregulation (CASP3/DR) in tumors frequently confers resistance to cancer therapy and is significantly correlated with a poor prognosis in cancer patients. Because CASP3/DR cancer cells rely heavily on the activity of caspase-7 (CASP7) to initiate apoptosis, inhibition of activated CASP7 (p19/p12-CASP7) by X-linked inhibitor of apoptosis protein (XIAP) is a potential mechanism by which apoptosis is prevented in those cancer cells. Here, we identify the pocket surrounding the Cys(246) residue of p19/p12-CASP7 as a target for the development of a protein-protein interaction (PPI) inhibitor of the XIAP:p19/p12-CASP7 complex. Interrupting this PPI directly triggered CASP7-dependent apoptotic signaling that bypassed the activation of the apical caspases and selectively killed CASP3/DR malignancies in vitro and in vivo without adverse side effects in nontumor cells. Importantly, CASP3/DR combined with p19/p12-CASP7 accumulation correlated with the aggressive evolution of clinical malignancies and a poor prognosis in cancer patients. Moreover, targeting of this PPI effectively killed cancer cells with multidrug resistance due to microRNA let-7a-1-mediated CASP3/DR and resensitized cancer cells to chemotherapy-induced apoptosis. These findings not only provide an opportunity to treat CASP3/DR malignancies by targeting the XIAP:p19/p12-CASP7 complex, but also elucidate the molecular mechanism underlying CASP3/DR in cancers.
The Size Effect on Solidification in Eutectic Bismuth-Tin (Bi-Sn) Nanowires by In-Situ Annealing Processes
(2010-10-01)
Chen, Shih-Hsun
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Wang, Chiu-Yen
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Chen, Lih-Juann
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Liu, Tzeng-Feng
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Chao, Chuen-Guang
The size effects on solidification and the formation mechanism of the segmented eutectic Bi-43Sn nanowires during in situ annealing have been investigated. A directional solidification along the wire axis limits the segmented eutectic nanowire to arrange axially during the in situ annealing processes due to directional solidification. In 70 nm nanowires, the small size confines the convection in liquid, which results in differences in the microstructure and composition profiles between 70 and 200 nm nanowires. In the vacuum hydraulic pressure injection process, the directional cooling helps the formation of single crystal, and the isotropic solidification leads to polycrystalline microstructure.