• Title/Summary/Keyword: ${\pi}$-domain

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Development of ELISA System for Screening of Specific Binding Inhibitors for Src Homology (SH)2 Domain and Phosphotyrosine Interactions

  • Lee, Sang-Seop;Lee, Kyung-Im;Yoo, Ji-Yun;Jeong, Moon-Jin;Park, Young-Mee;Kwon, Byoung-Mog;Bae, Yun-Soo;Han, Mi-Young
    • BMB Reports
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    • v.34 no.6
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    • pp.537-543
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    • 2001
  • In the present study, an in vitro ELISA system to assess the interaction between Src homology (SH)2 domains and phosphotyrosine that contain peptides was established using purified GST-conjugated SH2 proteins and synthetic biotinylated phosphotyrosine that contain oligopeptides. The SH2 domains bound the relevant phosphopeptides that were immobilized in the streptavidin-coated microtiter plate in a highly specific and dose-dependent manner. The epidermal growth factor receptor (EGFR)-, T antigen (T Ag)-, and platelet-derived growth factor receptor (PDGFR)-derived phosphopeptides interacted with the growth factor receptor binding protein (Grb)2/SH2, Lck/SH2, and phosphatidyl inositol 3-kinase (PI3K) p85/SH2, respectively. No cross-reactions were observed. Competitive inhibition experiments showed that a short phosphopeptide of only four amino acids was long enough to determine the binding specificity. Optimal concentrations of the GST-SH2 fusion protein and phosphopeptide in this new ELISA system for screening the binding blockers were chosen at 2nM and 500nM, respectively. When two candidate compounds were tested in our ELISA system, they specifically inhibited the Lck/SH2 and/or p85/SH2 binding to the relevant phosphopeptides. Our results indicate that this ELISA system could be used as an easy screening method for the discovery of specific binding blockers of protein-protein interactions via SH2 domains.

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A Novel Simple Method to Purify Recombinant Soluble Human Complement Receptor Type 1 (sCR 1) from CHO Cell Culture

  • Wang, Pi-Chao;Hisamune Kato;Takehiro Inoue;Masatoshi Matsumura;Noriyuki Ishii;Yoshinobu Murakami;Tsukasa Seya
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.2
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    • pp.67-75
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    • 2002
  • The human complement receptor type 1 (CR 1, C3 b/C4b receptor) is a polymorphic membrane glycoprotein expressed on human erythrocytes, peripheral leukocytes, plasma and renal glomerular podocytes, which consists of transmembrane and cytoplasmic domains with 30 repeating homologous protein domains known as short consensus repeats (SCR). CR1 has been used as an inhibitor for inflammatory and immune system for the past several years. Recently; it is reported that CRl was found to suppress the hyper-acute rejection in xeno-transplantation and can be used to cure autoimmune diseases. A soluble form of CRl, called sCRl, is a recombinant CRl by cleaving the transmembrane domain at C-terminus and has been expressed in Chinese Hamster Ovary (CHO) cells. Several purification methods for sCR1 from CHO cells have been reported, but most of them require complicated steps at high cost. Moreover, such methods are mostly performed under the pH condition apt to denaturing sCR1 and causes sCRl losing its activity. We here report a rapid and efficient method to purify sCR1 from CHO cell. The new method consists of a two-stage of cell culture by cultivating cells in serum medium followed by serum-free medium, and a two-stage of column purification by means of heparin and gel filtration column chromatography. By using this novel method, sCR1 can be purified in a simple and effective way with high yield and purity, furthermore, the purified sCR1 was confirmed to retain its activity to suppress the complement activation in vivo and ex vivo.

VIRTUAL PREDICTION OF A RADIAL-PLY TIRE'S IN-PLANE FREE VIBRATION MODES TRANSMISSIBILITY

  • CHANG Y. P.;EL-GINDY M.
    • International Journal of Automotive Technology
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    • v.6 no.2
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    • pp.149-159
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    • 2005
  • A full nonlinear finite element P185/70Rl4 passenger car radial-ply tire model was developed and run on a 1.7-meter-diameter spinning test drum/cleat model at a constant speed of 50 km/h in order to investigate the tire transient response characteristics, i.e. the tire in-plane free vibration modes transmissibility. The virtual tire/drum finite element model was constructed and tested using the nonlinear finite element analysis software, PAM-SHOCK, a nonlinear finite element analysis code. The tire model was constructed in extreme detail with three-dimensional solid, layered membrane, and beam finite elements, incorporating over 18,000 nodes and 24 different types of materials. The reaction forces of the tire axle in vertical (Z axis) and longitudinal (X axis) directions were recorded when the tire rolled over a cleat on the drum, and then the FFT algorithm was applied to examine the transient response information in the frequency domain. The result showed that this PI 85/70Rl4 tire has clear peaks of 84 and 45 Hz transmissibility in the vertical and longitudinal directions. This result was validated against more than 10 previous studies by either theoretical or experimental approaches and showed excellent agreement. The tire's post-impact response was also investigated to verify the numerical convergence and computational stability of this FEA tire model and simulation strategy, the extraordinarily stable scenario was confirmed. The tire in-plane free vibration modes transmissibility was successfully detected. This approach was never before attempted in investigations of tire in-plane free vibration modes transmission phenomena; this work is believed to be the first of its kind.

ATAD2 expression increases [18F]Fluorodeoxyglucose uptake value in lung adenocarcinoma via AKT-GLUT1/HK2 pathway

  • Sun, Tong;Du, Bulin;Diao, Yao;Li, Xuena;Chen, Song;Li, Yaming
    • BMB Reports
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    • v.52 no.7
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    • pp.457-462
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    • 2019
  • [18F]Fluorodeoxyglucose (FDG) PET/CT imaging has been widely used in the diagnosis of malignant tumors. ATPase family AAA domain-containing protein 2 (ATAD2) plays important roles in tumor growth, invasion and metastasis. However, the relationship between [18F]FDG accumulation and ATAD2 expression remains largely unknown. This study aimed to investigate the correlation between ATAD2 expression and [18F]FDG uptake in lung adenocarcinoma (LUAD), and elucidate its underlying molecular mechanisms. The results showed that ATAD2 expression was positively correlated with maximum standardized uptake value ($SUV_{max}$), total lesion glycolysis (TLG), glucose transporter type 1 (GLUT1) expression and hexokinase2 (HK2) expression in LUAD tissues. In addition, ATAD2 knockdown significantly inhibited the proliferation, tumorigenicity, migration, [18F]FDG uptake and lactate production of LUAD cells, while, ATAD2 overexpression exhibited the opposite effects. Furthermore, ATAD2 modulated the glycometabolism of LUAD via AKT-GLUT1/HK2 pathway, as assessed using LY294002 (an inhibitor of PI3K/AKT pathway). In summary, to explore the correlation between ATAD2 expression and glycometabolism is expected to bring good news for anti-energy metabolism therapy of cancers.

Stage specific transcriptome profiles at cardiac lineage commitment during cardiomyocyte differentiation from mouse and human pluripotent stem cells

  • Cho, Sung Woo;Kim, Hyoung Kyu;Sung, Ji Hee;Han, Jin
    • BMB Reports
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    • v.54 no.9
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    • pp.464-469
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    • 2021
  • Cardiomyocyte differentiation occurs through complex and finely regulated processes including cardiac lineage commitment and maturation from pluripotent stem cells (PSCs). To gain some insight into the genome-wide characteristics of cardiac lineage commitment, we performed transcriptome analysis on both mouse embryonic stem cells (mESCs) and human induced PSCs (hiPSCs) at specific stages of cardiomyocyte differentiation. Specifically, the gene expression profiles and the protein-protein interaction networks of the mESC-derived platelet-derived growth factor receptor-alpha (PDGFRα)+ cardiac lineage-committed cells (CLCs) and hiPSC-derived kinase insert domain receptor (KDR)+ and PDGFRα+ cardiac progenitor cells (CPCs) at cardiac lineage commitment were compared with those of mesodermal cells and differentiated cardiomyocytes. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses revealed that the genes significantly upregulated at cardiac lineage commitment were associated with responses to organic substances and external stimuli, extracellular and myocardial contractile components, receptor binding, gated channel activity, PI3K-AKT signaling, and cardiac hypertrophy and dilation pathways. Protein-protein interaction network analysis revealed that the expression levels of genes that regulate cardiac maturation, heart contraction, and calcium handling showed a consistent increase during cardiac differentiation; however, the expression levels of genes that regulate cell differentiation and multicellular organism development decreased at the cardiac maturation stage following lineage commitment. Additionally, we identified for the first time the protein-protein interaction network connecting cardiac development, the immune system, and metabolism during cardiac lineage commitment in both mESC-derived PDGFRα+ CLCs and hiPSC-derived KDR+PDGFRα+ CPCs. These findings shed light on the regulation of cardiac lineage commitment and the pathogenesis of cardiometabolic diseases.

RINGS WITH A RIGHT DUO FACTOR RING BY AN IDEAL CONTAINED IN THE CENTER

  • Cheon, Jeoung Soo;Kwak, Tai Keun;Lee, Yang;Piao, Zhelin;Yun, Sang Jo
    • Bulletin of the Korean Mathematical Society
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    • v.59 no.3
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    • pp.529-545
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    • 2022
  • This article concerns a ring property that arises from combining one-sided duo factor rings and centers. A ring R is called right CIFD if R/I is right duo by some proper ideal I of R such that I is contained in the center of R. We first see that this property is seated between right duo and right π-duo, and not left-right symmetric. We prove, for a right CIFD ring R, that W(R) coincides with the set of all nilpotent elements of R; that R/P is a right duo domain for every minimal prime ideal P of R; that R/W(R) is strongly right bounded; and that every prime ideal of R is maximal if and only if R/W(R) is strongly regular, where W(R) is the Wedderburn radical of R. It is also proved that a ring R is commutative if and only if D3(R) is right CIFD, where D3(R) is the ring of 3 by 3 upper triangular matrices over R whose diagonals are equal. Furthermore, we show that the right CIFD property does not pass to polynomial rings, and that the polynomial ring over a ring R is right CIFD if and only if R/I is commutative by a proper ideal I of R contained in the center of R.

A Study of Textured Image Segmentation using Phase Information (페이즈 정보를 이용한 텍스처 영상 분할 연구)

  • Oh, Suk
    • Journal of the Korea Society of Computer and Information
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    • v.16 no.2
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    • pp.249-256
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    • 2011
  • Finding a new set of features representing textured images is one of the most important studies in textured image analysis. This is because it is impossible to construct a perfect set of features representing every textured image, and it is inevitable to choose some relevant features which are efficient to on-going image processing jobs. This paper intends to find relevant features which are efficient to textured image segmentation. In this regards, this paper presents a different method for the segmentation of textured images based on the Gabor filter. Gabor filter is known to be a very efficient and effective tool which represents human visual system for texture analysis. Filtering a real-valued input image by the Gabor filter results in complex-valued output data defined in the spatial frequency domain. This complex value, as usual, gives the module and the phase. This paper focused its attention on the phase information, rather than the module information. In fact, the module information is considered very useful at region analysis in texture, while the phase information was considered almost of no use. But this paper shows that the phase information can also be fully useful and effective at region analysis in texture, once a good method introduced. We now propose "phase derivated method", which is an efficient and effective way to compute the useful phase information directly from the filtered value. This new method reduces effectively computing burden and widen applicable textured images.

Physiological Roles of Phospholipase Cγ and Its Mutations in Human Disease (Phospholipase Cγ의 생리적 기능과 질병과 연관된 돌연변이)

  • Jang, Hyun-Jun;Choi, Jang Hyun;Chang, Jong-Soo
    • Journal of Life Science
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    • v.30 no.9
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    • pp.826-833
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    • 2020
  • Phospholipase C gamma (PLCγ) has critical roles in receptor tyrosine kinase- and non-receptor tyrosine kinase-mediated cellular signaling relating to the hydrolysis of phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] to produce inositol 1,4,5 trisphosphate (IP3) and diacylglycerol (DAG), which promote protein kinase C (PKC) and Ca2+ signaling to their downstream cellular targets. PLCγ has two isozymes called PLCγ1 and PLCγ2, which control cell growth and differentiation. In addition to catalytically active X- and Y-domains, both isotypes contain two Src homology 2 (SH2) domains and an SH3 domain for protein-protein interaction when the cells are activated by ligand stimulation. PLCγ also contains two pleckstrin homology (PH) domains for membrane-associated phosphoinositide binding and protein-protein interactions. While PLCγ1 is widely expressed and appears to regulate intracellular signaling in many tissues, PLCγ2 expression is restricted to cells of hematopoietic systems and seems to play a role in the regulation of immune response. A distinct mechanism for PLCγ activation is linked to an increase in phosphorylation of specific tyrosine residue, Y783. Recent studies have demonstrated that PLCγ mutations are closely related to cancer, immune disease, and brain disorders. Our review focused on the physiological roles of PLCγ by means of its structure and enzyme activity and the pathological functions of PLCγ via mutational analysis obtained from various human diseases and PLCγ knockout mice.

Molecular Cloning and Characterization of a Novel Stem-specific Gene from Camptotheca acuminata

  • Pi, Yan;Liao, Zhihua;Chai, Yourong;Zeng, Hainian;Wang, Peng;Gong, Yifu;Pang, Yongzhen;Sun, Xiaofen;Tang, Kexuan
    • BMB Reports
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    • v.39 no.1
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    • pp.68-75
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    • 2006
  • In higher plants, P450s participate in the biosynthesis of many important secondary metabolites. Here we reported for the first time the isolation of a new cytochrome P450 cDNA that expressed in a stem-specific manner from Camptotheca acuminata (designated as CaSS), a native medicinal plant species in China, using RACE-PCR. The full-length cDNA of CaSS was 1735 bp long containing a 1530 bp open reading frame (ORF) encoding a polypeptide of 509 amino acids. Bioinformatic analysis revealed that CASS contained a heme-binding domain PFGXGRRXCX and showed homology to other plant cytochrome P450 monooxygenases and hydroxylases. Southern blotting analysis revealed that there was only one copy of the CaSS present in the genome of Camptotheca acuminata. Northern blotting analysis revealed that CaSS expressed, in a tissue-specific manner, highly in stem and lowly in root, leaf and flower. Our study suggests that CaSS is likely to be involved in the phenylpropanoid pathway.

A Historical and Mathematical Analysis on the Radian (라디안 개념의 역사적 분석과 수학적 분석)

  • Yoo, Jaegeun;Lee, Kyeong-Hwa
    • Journal of Educational Research in Mathematics
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    • v.27 no.4
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    • pp.833-855
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    • 2017
  • This study aims to reinvestigate the reason for introducing radian as a new unit to express the size of angles, what is the meaning of radian measures to use arc lengths as angle measures, and why is the domain of trigonometric functions expanded to real numbers for expressing general angles. For this purpose, it was conducted historical, mathematical and applied mathematical analyzes in order to research at multidisciplinary analysis of the radian concept. As a result, the following were revealed. First, radian measure is intrinsic essence in angle measure. The radian is itself, and theoretical absolute unit. The radian makes trigonometric functions as real functions. Second, radians should be aware of invariance through covariance of ratios and proportions in concentric circles. The orthogonality between cosine and sine gives a crucial inevitability to the radian. It should be aware that radian is the simplest standards for measuring the length of arcs by the length of radius. It can find the connection with sexadecimal method using the division strategy. Third, I revealed the necessity by distinction between angle and angle measure. It needs justification for omission of radians and multiplication relationship strategy between arc and radius. The didactical suggestions derived by these can reveal the usefulness and value of the radian concept and can contribute to the substantive teaching of radian measure.