• Title/Summary/Keyword: domain expression

Search Result 898, Processing Time 0.028 seconds

Expression, Refolding, and Characterization of the Proteolytic Domain of Human Bone Morphogenetic Protein 1 (뼈형성 단백질(Bone Morphogenetic Protein 1)의 단백질 분해 부위의 발현 및 특성 연구)

  • ;Daihung Do
    • Journal of Life Science
    • /
    • v.10 no.2
    • /
    • pp.218-227
    • /
    • 2000
  • Bone morphogenetic protein 1 (BMP-1) is part of a complex capable of inducing ectopic bone formation in mammals. Studies on TGF-β1 processing and Drosophila dorsal-ventral patterning have focused attention on BMP-1 as important in mediating the biological activity of this bone inducing complex. Herein, the bacterial expression, refolding, purification, and initial characterization of the BMP-1 proteolytic domain (BPD) are described. A semi-quantitative fluorescence-based thin layer chromatography assay was developed to assist in rapidly screening for optimal renaturation conditions. According to a preliminary screen for optimal conditions for the refolding of BPD , a detectable proteolytic activity against a high turnover substrate for astacin, a homologous protease from crayfish was observed. The conditions identified have allowed the expression of sufficient amounts of BPD for the characterization of the protein. Its proteolytic activity exhibits the same cleavage specificity as astacin against seven substrates that were previously synthesized for studying astacin. Furthermore, this activity is inhibited by the metal chelator 1,10-phenanthroline but not by its analogue 1,7-phenanthroline. The collagenase inhibitor Pro-Leu-Gly hydroxamate was found to inhibit both astacin and BPD activity. The results presented in this paper argue that BMP-1 does in fact possess an intrinsic proteolytic activity.

  • PDF

Efficacy of Gene Transfer and Expression of Novel Recombinant Baculovirus Vector (새로운 재조합 베큘로바이러스 벡터의 유전자 전달과 유전자 발현의 효과)

  • Kweon, Tae-Dong;Hong, Seong-Karp
    • Journal of the Korea Institute of Information and Communication Engineering
    • /
    • v.18 no.8
    • /
    • pp.2017-2022
    • /
    • 2014
  • Novel baculovirus vector systems recombined with coding genes of polyhedron promoter, vesicular stomatitis virus G (VSVG), polyA, cytomegalovirus (CMV) promoter, enhanced green fluorescent protein (EGFP), and protein transduction domain (PTD) were constructed. These recombinant baculovirus vector systems were applied into human foreskin fibroblast cells and compared the effects of gene transfer and gene expression of these recombinant baculovirus vector systems with control vector system. From this study, it showed that these novel recombinant baculovirus vector systems were superior efficacy to control vector system in view of gene transfer and gene expression.

Expression of Recombinant Rotavirus Proteins Harboring Antigenic Epitopes of the Hepatitis A Virus Polyprotein in Insect Cells

  • Than, Van Thai;Baek, In-Hyuk;Lee, Hee-Young;Kim, Jong-Bum;Shon, Dong-Hwa;Chung, In-Sik;Kim, Won-Yong
    • Biomolecules & Therapeutics
    • /
    • v.20 no.3
    • /
    • pp.320-325
    • /
    • 2012
  • Rotavirus and hepatitis A virus (HAV) spread by the fecal-oral route and infections are important in public health, especially in developing countries. Here, two antigenic epitopes of the HAV polyprotein, domain 2 (D2) and domain 3 (D3), were recombined with rotavirus VP7, generating D2/VP7 and D3/VP7, cloned in a baculovirus expression system, and expressed in Spodoptera frugiperda 9 (Sf9) insect cells. All were highly expressed, with peak expression 2 days post-infection. Western blotting and ELISA revealed that two chimeric proteins were antigenic, but only D2/VP7 was immunogenic and elicited neutralizing antibody responses against rotavirus and HAV by neutralization assay, implicating D2/VP7 as a multivalent subunit-vaccine Candidate for preventing both rotavirus and HAV infections.

Overexpression of FGFR3 mRNA and Mutational Analysis of FGFR3 Gene in Hepatocellular Carcinoma (간암에서 FGFR3 mRNA의 과발현과 FGFR3 유전자의 돌연변이 분석)

  • Chang, Young Gyoon;Bae, Hyun Jin;Nam, Suk Woo
    • YAKHAK HOEJI
    • /
    • v.56 no.6
    • /
    • pp.352-357
    • /
    • 2012
  • FGFR3 is a member of the fibroblast growth factor receptor family which interacts with fibroblast growth factors, setting in motion a cascade of downstream signals, ultimately influencing mitogenesis and differentiation. This particular family member binds acidic and basic fibroblast growth hormone and plays a role in bone development and maintenance. Accumulated evidence suggests that aberrant regulation of FGFR3 and genetic alterations are implicated in the development and progression of various cancers. Despite a high incidence of FGFR3 over-expression, no such investigation has been performed in hepatocellular carcinoma. Thus, we investigated genetic alterations of the FGFR3 gene in 73 cases of hepatocellular carcinoma by single-strand conformational polymorphism (SSCP) and sequencing. One silent mutation (A369A) was found in the extracellular domain of FGFR3, and one genetic alteration in the immunoglobulin-like III domain of FGFR3 appeared to be polymorphism. Taken together, we concluded that over-expression of FGFR3 in hepatocellular carcinoma is not associated with genetic alterations of FGFR3 gene, and we suggest that there could be another underlying mechanism of aberrant FGFR3 expression in hepatocellular carcinoma.

Genome-wide analysis of Solanum lycopersicum L. cyclophilins

  • Khatun, Khadiza;Robin, Arif Hasan Khan;Islam, Md. Rafiqul;Jyoti, Subroto Das;Lee, Do-Jin;Kim, Chang Kil;Chung, Mi-Young
    • Journal of Plant Biotechnology
    • /
    • v.49 no.1
    • /
    • pp.15-29
    • /
    • 2022
  • Cyclophilins (CYPs) are highly conserved ubiquitous proteins belong to the peptidyl prolyl cis/trans isomerase (PPIase) superfamily. These proteins are present in a wide range of organisms; they contain a highly conserved peptidyl-prolyl cis/trans isomerase domain. A comprehensive database survey identified a total of 35 genes localized in all cellular compartments of Solanum lycopersicum L., but largely in the cytosol. Sequence alignment and conserved motif analyses of the SlCYP proteins revealed a highly conserved CLD motif. Evolutionary analysis predicted the clustering of a large number of gene pairs with high sequence similarity. Expression analysis using the RNA-Seq data showed that the majority of the SlCYP genes were highly expressed in mature leaves and blooming flowers, compared with their expression in other organs. This study provides a basis for the functional characterization of individual CYP genes in the future to elucidate their role(s) in protein refolding and long-distance signaling in tomatoes and in plant biology, in general.

Image Compression by Texture Expression Method of Wavelet Coefficients (웨이브렛 계수의 텍스춰 표현에 의한 영상 압축)

  • Wang, Jiang-Qing;Park, Min-Sheik;Kwak, Hoon-Sung
    • Journal of the Institute of Electronics Engineers of Korea TE
    • /
    • v.39 no.3
    • /
    • pp.83-89
    • /
    • 2002
  • A new scheme for image compression based on texture expression in the wavelet transform domain is presented. After taking wavelet transform, using the fact that the high-pass filtered bands has a lower variance than that of the original, a texture expression for the homogeneous polygonal regions can be more efficiently performed in the wavelet transform domain. The estimated texture parameters are transmitted to the receiver and later used for reconstruction after storing in disk. In most cases, the proposed method has yields good results with respects to the compression ratio and reconstructed image quality when our system has compared to conventional SPIHT scheme. 

Secondary Structure, 1H, 13C and 15N Resonance Assignments and Molecular Interactions of the Dishevelled DIX Domain

  • Capelluto, Daniel G.S.;Overduin, Michael
    • BMB Reports
    • /
    • v.38 no.2
    • /
    • pp.243-247
    • /
    • 2005
  • Dishevelled (Dvl) is a positive regulator of the canonical Wnt signaling pathway, which regulates the levels of $\beta$-catenin. The $\beta$-catenin oncoprotein depends upon the association of Dvl and Axin proteins through their DIX domains, and its accumulation directs the expression of specific developmental-related genes at the nucleus. Here, the $^1H$, $^{13}C$, and $^{15}N$ resonances of the human Dishevelled 2 DIX domain are assigned using heteronuclear nuclear magnetic resonance (NMR) spectroscopy. In addition, helical and extended elements are identified based on the NMR data. The results establish a structural context for characterizing the actin and phospholipid interactions and binding sites of this novel domain, and provide insights into its role in protein localization to stress fibers and cytoplasmic vesicles during Wnt signaling.

Characterization of tTA and Its Functional Domain in Tetracycline Repressor-mediated Gene Repression System

  • Kim, Hong-Jin;Kim, Ki-Ho
    • Archives of Pharmacal Research
    • /
    • v.21 no.3
    • /
    • pp.320-325
    • /
    • 1998
  • To elucidate of role(s) of tTA as a repressor in the tTA-mediated gene repression system, we introduced mutations into the acidic domain of VP16 and examined the effects of such various mutations. In the transient repression experiment, a region containing 34 amino acids of the activation domain of VP16 (412-456) which interacts with TFIIB was found to be necessary and sufficient for the tTA-mediated repression of gene expression. However, in the experiment to investigate the fact that tTA-regulated repression is related to the activation function of VP16, we found that the repression abilities of tTA derivatives did not correlate exactly with their activation abilities. Therefore, we conclude that increased mass of VP16 in tTA might be also important for efficient repression in addition to functional domain of VP16.

  • PDF

A Frequency Domain Equalization Algorithm for Fast Time-Varying Fading Channels

  • Tran, Le-Nam;Hong, Een-Kee;Liu, Huaping
    • Journal of Communications and Networks
    • /
    • v.11 no.5
    • /
    • pp.473-479
    • /
    • 2009
  • Conventional frequency domain equalization (FDE) schemes were originally devised for quasi-static channels. Thus, such equalization schemes could suffer from significant performance degradation in fast-fading channels. This paper proposes a frequency domain equalization algorithm to mitigate the effect of fast time-varying fading. First, a mathematical expression is derived to quantify the total interference resulting from the time variation of the channel. Then, the proposed approach attempts to eliminate the effect of time-variations of the channel. This cancellation allows efficient use of the classical FDE structures in fast time-varying fading environments, although they are built upon the quasi-static channel model. Simulation results of bit-error-rate performance are provided to demonstrate the effectiveness of the proposed algorithm.

Pleckstrin homology domain of phospholipase D2 is a negative regulator of focal adhesion kinase

  • Kim, Mi Kyoung;Hwang, Won Chan;Min, Do Sik
    • BMB Reports
    • /
    • v.54 no.2
    • /
    • pp.112-117
    • /
    • 2021
  • Phospholipase D2 (PLD2) has been implicated in the tyrosine kinase-mediated signaling pathways, but the regulation events are yet to be identified. Herein, we demonstrate that pleckstrin homology (PH) domain of PLD2 (PLD2-PH) exerts an antitumorigenic effect via the suppression of PLD2 and focal adhesion kinase (FAK). The kinase domain of FAK interacts with PLD2-PH and induces tyrosine phosphorylation and activation of PLD2. Furthermore, PLD2 increased tyrosine phosphorylation of FAK. However, ectopic expression of the PLD2-PH competes for binding to FAK and reduces the interaction between PLD2 and FAK, thereby suppressing FAK-induced PLD activation and tyrosine phosphorylation of FAK. The PLD2-PH suppressed the migration and invasion of glioblastoma cells, as well as tumor formation in a xenograft mouse model. This study uncovers a novel role of PLD2-PH as a negative regulator of PLD2 and FAK.