• Title/Summary/Keyword: terminal region

Search Result 612, Processing Time 0.023 seconds

Structure-activity relationships of cecropin-like peptides and their interactions with phospholipid membrane

  • Lee, Eunjung;Jeong, Ki-Woong;Lee, Juho;Shin, Areum;Kim, Jin-Kyoung;Lee, Juneyoung;Lee, Dong Gun;Kim, Yangmee
    • BMB Reports
    • /
    • v.46 no.5
    • /
    • pp.282-287
    • /
    • 2013
  • Cecropin A and papiliocin are novel 37-residue cecropin-like antimicrobial peptides isolated from insect. We have confirmed that papiliocin possess high bacterial cell selectivity and has an ${\alpha}$-helical structure from $Lys^3$ to $Lys^{21}$ and from $Ala^{25}$ to $Val^{35}$, linked by a hinge region. In this study, we demonstrated that both peptides showed high antimicrobial activities against multi-drug resistant Gram negative bacteria as well as fungi. Interactions between these cecropin-like peptides and phospholipid membrane were studied using CD, dye leakage experiments, and NMR experiments, showing that both peptides have strong permeabilizing activities against bacterial cell membranes and fungal membranes as well as $Trp^2$ and $Phe^5$ at the N-terminal helix play an important role in attracting cecropin-like peptides to the negatively charged bacterial cell membrane. Cecropin-like peptides can be potent peptide antibiotics against multi-drug resistant Gram negative bacteria and fungi.

Antimicrobial Activity of Bacteriophage Endolysin Produced in Nicotiana benthamiana Plants

  • Kovalskaya, Natalia;Foster-Frey, Juli;Donovan, David M.;Bauchan, Gary;Hammond, Rosemarie W.
    • Journal of Microbiology and Biotechnology
    • /
    • v.26 no.1
    • /
    • pp.160-170
    • /
    • 2016
  • The increasing spread of antibiotic-resistant pathogens has raised the interest in alternative antimicrobial treatments. In our study, the functionally active gram-negative bacterium bacteriophage CP933 endolysin was produced in Nicotiana benthamiana plants by a combination of transient expression and vacuole targeting strategies, and its antimicrobial activity was investigated. Expression of the cp933 gene in E. coli led to growth inhibition and lysis of the host cells or production of trace amounts of CP933. Cytoplasmic expression of the cp933 gene in plants using Potato virus X-based transient expression vectors (pP2C2S and pGR107) resulted in death of the apical portion of experimental plants. To protect plants against the toxic effects of the CP933 protein, the cp933 coding region was fused at its Nterminus to an N-terminal signal peptide from the potato proteinase inhibitor I to direct CP933 to the delta-type vacuoles. Plants producing the CP933 fusion protein did not exhibit the severe toxic effects seen with the unfused protein and the level of expression was 0.16 mg/g of plant tissue. Antimicrobial assays revealed that, in contrast to gram-negative bacterium E. coli (BL21(DE3)), the gram-positive plant pathogenic bacterium Clavibacter michiganensis was more susceptible to the plant-produced CP933, showing 18% growth inhibition. The results of our experiments demonstrate that the combination of transient expression and protein targeting to the delta vacuoles is a promising approach to produce functionally active proteins that exhibit toxicity when expressed in plant cells.

Expression of Hr-Erf Gene during Ascidian Embryogenesis

  • Kim, Jung Eun;Lee, Won Young;Kim, Gil Jung
    • Development and Reproduction
    • /
    • v.17 no.4
    • /
    • pp.389-397
    • /
    • 2013
  • FGF9/16/20 signaling pathway specify the developmental fates of notochord, mesenchyme, and neural cells in ascidian embryos. Although a conserved Ras/MEK/Erk/Ets pathway is known to be involved in this signaling, the detailed mechanisms of regulation of FGF signaling pathway have remained largely elusive. In this study, we have isolated Hr-Erf, an ascidian orthologue of vertebrate Erf, to elucidate interactions of transcription factors involved in FGF signaling of the ascidian embryo. The Hr-Erf cDNA encompassed 3110 nucleotides including sequence encoded a predicted polypeptide of 760 amino acids. The polypeptide had the Ets DNA-binding domain in its N-terminal region. In adult animals, Hr-Erf mRNA was predominantly detected in muscle, and at lower levels in ganglion, gills, gonad, hepatopancreas, and stomach by quantitative real-time PCR (QPCR) method. During embryogenesis, Hr-Erf mRNA was detected from eggs to early developmental stage embryos, whereas the transcript levels were decreased after neurula stage. Similar to the QPCR results, maternal transcripts of Hr-Erf was detected in the fertilized eggs by whole-mount in situ hybridization. Maternal mRNA of Hr-Erf was gradually lost from the neurula stage. Zygotic expression of Hr-Erf started in most blastomeres at the 8-cell stage. At gastrula stage, Hr-Erf was specifically expressed in the precursor cells of brain and mesenchyme. When MEK inhibitor was treated, embryos resulted in loss of Hr-Erf expression in mesenchyme cells, and in excess of Hr-Erf in a-line neural cells. These results suggest that zygotic Hr-Erf products are involved in specification of mesenchyme and neural cells.

Efficient Contour Coding for Segmentation-Based Image Coding (분할 기반 영상 부호화를 위한 효율적 윤곽선 부호화)

  • Kim, Gi-Seok;Park, Young-Sik;Song, Kun-Woen;Chung, Eui-Yoon;Kim, Yong-Suk;Ha, Yeong-Ho
    • Journal of the Korean Institute of Telematics and Electronics S
    • /
    • v.35S no.11
    • /
    • pp.152-165
    • /
    • 1998
  • The contour coding usually occupies the biggest part in the encoded bitstream, which causes the bottleneck problem of a region-based coding scheme. In this paper, new adaptive contour coding technique is proposed for the segmentation-based image coding. By adaptive contour coding considering contrast of neighbor regions in the proposed method, the overall bitrate can be significantly reduced without loss of the subjective image quality. After segmentation using watershed algorithm to the image, the contour segments are classified according to the contrast of the adjacent regions. Then, the contour segments between classified low contrast regions are highly compressed using morphological low pass filtering. The needed bits for encoding the contour information is reduced without loss of subjective image quality in the experiment.

  • PDF

Correlates of Digit Bias in Self-reporting of Cigarette per Day (CPD) Frequency: Results from Global Adult Tobacco Survey (GATS), India and its Implications

  • Jena, Pratap Kumar;Kishore, Jugal;Jahnavi, G.
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.14 no.6
    • /
    • pp.3865-3869
    • /
    • 2013
  • Background: Cigarette per day (CPD) use is a key smoking behaviour indicator. It reflects smoking intensity which is directly proportional to the occurrence of tobacco induced cancers. Self reported CPD assessment in surveys may suffer from digit bias and under reporting. Estimates from such surveys could influence the policy decision for tobacco control efforts. In this context, this study aimed at identifying underlying factors of digit bias and its implications for Global Adult Tobacco Surveillance. Materials or Methods: Daily manufactured cigarette users CPD frequencies from Global Adult Tobacco Survey (GATS) - India data were analyzed. Adapted Whipple Index was estimated to assess digit bias and data quality of reported CPD frequency. Digit bias was quantified by considering reporting of '0' or '5' as the terminal digits in the CPD frequency. The factors influencing it were identified by bivariate and logistic regression analysis. Results: The mean and mode of CPD frequency was 6.7 and 10 respectively. Around 14.5%, 15.1% and 15.2% of daily smokers had reported their CPD frequency as 2, 5 and 10 respectively. Modified Whipple index was estimated to be 226.3 indicating poor data quality. Digit bias was observed in 38% of the daily smokers. Heavy smoking, urban residence, North, South, North- East region of India, less than primary, secondary or higher educated and fourth asset index quintile group were significantly associated with digit bias. Discussion: The present study highlighted poor quality of CPD frequency data in the GATS-India survey and need for its improvement. Modeling of digit preference and smoothing of the CPD frequency data is required to improve quality of data. Marketing of 10 cigarette sticks per pack may influence CPD frequency reporting, but this needs further examination. Exploring alternative methods to reduce digit bias in cross sectional surveys should be given priority.

JQ1, a BET inhibitor, controls TLR4-induced IL-10 production in regulatory B cells by BRD4-NF-κB axis

  • Lee, Min Bum;Lee, Jun-Ho;Hong, Seong Hwi;You, Jueng Soo;Nam, Seung Taek;Kim, Hyun Woo;Park, Young Hwan;Lee, Dajeong;Min, Keun Young;Park, Yeong-Min;Kim, Young Mi;Kim, Hyuk Soon;Choi, Wahn Soo
    • BMB Reports
    • /
    • v.50 no.12
    • /
    • pp.640-646
    • /
    • 2017
  • Regulatory B cells, also well-known as IL-10-producing B cells, play a role in the suppression of inflammatory responses. However, the epigenetic modulation of regulatory B cells is largely unknown. Recent studies showed that the bromodomain and extra-terminal domain (BET) protein inhibitor JQ1 controls the expression of various genes involving cell proliferation and cell cycle. However, the role of BET proteins on development of regulatory B cells is not reported. In this study, JQ1 potently suppressed IL-10 expression and secretion in murine splenic and peritoneal B cells. While bromodomain-containing protein 4 (BRD4) was associated with $NF-{\kappa}B$ on IL-10 promoter region by LPS stimulation, JQ1 interfered the interaction of BRD4 with $NF-{\kappa}B$ on IL-10 promoter. In summary, BRD4 is essential for toll like receptor 4 (TLR4)-mediated IL-10 expression, suggesting JQ1 could be a potential candidate in regulating IL-10-producing regulatory B cells in cancer.

Localization of Barley yellow dwarf virus Movement Protein Modulating Programmed Cell Death in Nicotiana benthamiana

  • Ju, Jiwon;Kim, Kangmin;Lee, Kui-Jae;Lee, Wang Hu;Ju, Ho-Jong
    • The Plant Pathology Journal
    • /
    • v.33 no.1
    • /
    • pp.53-65
    • /
    • 2017
  • Barley yellow dwarf virus (BYDV) belongs to Luteovirus and is limited only at phloem related tissues. An open reading frame (ORF) 4 of BYDV codes for the movement protein (MP) of BYDV gating plasmodesmata (PD) to facilitate virus movement. Like other Luteoviruses, ORF 4 of BYDV is embedded in the ORF3 but expressed from the different reading frame in leaky scanning manner. Although MP is a very important protein for systemic infection of BYDV, there was a little information. In this study, MP was characterized in terms of subcellular localization and programmed cell death (PCD). Gene of MP or its mutant (ΔMP) was expressed by Agroinfiltration method. MP was clearly localized at the nucleus and the PD, but ΔMP which was deleted distal N-terminus of MP showed no localization to PD exhibited the different target with original MP. In addition to PD localization, MP appeared associated with small granules in cytoplasm whereas ΔMP did not. MP associated with PD and small granules induced PCD, but ΔMP showed no association with PD and small granules did not exhibit PCD. Based on this study, the distal N-terminal region within MP is seemingly responsible for the localization of PD and the induction small granules and PCD induction. These results suggest that subcellular localization of BYDV MP may modulate the PCD in Nicotiana benthamiana.

A Study on the Revitalization of Yangsan ICD (양산 ICD 활성화 방안에 관한 연구)

  • Lee, Dong-Hun;Kim, Yul-Seong;Park, Ho;Shin, Jae-Yeong
    • Journal of Korea Port Economic Association
    • /
    • v.31 no.4
    • /
    • pp.121-132
    • /
    • 2015
  • Yangsan ICD (Inland Container Depot) has played an important role for Busan Port and Korea's port & logistics industries, increasing international logistics competitiveness and containers' transportation competitiveness in Busan region dealing with 1330 thousand TEU in 2005, since its opening in March 2000. However, it is necessary to seek new measures to revitalize Yangsan ICD, since its cargo volume decreased rapidly owing to the opening of Busan New Port and hinterland in 2006. This study constructed an evaluation model using AHP (Analytic Hierarchy Process) and conducted a survey targeting local businesses and persons concerned in Yangsan ICD to seek measures for revitalization. The results suggest that Yangsan ICD needs to switch functions to logistics centers (terminal facilities, logistics warehouse) for revitalization considering its advantage of facility location. Moreover, by extending the utilization period and securing building-to-land ratio, existing and new businesses' stable activity should be guaranteed. Furthermore, utilizing facilities such as the railway station in ICD, an active railway revitalization policy may increase cargo volume. Yangsan ICD should perform its role as an inland logistics depot through the revitalization of railway freight transportation in the national logistics system focusing on road freight transportation.

Choristoneura fumiferana Granulovirus p74 Protein, a Highly Conserved Baculoviral Envelope Protein

  • Rashidan, Kianoush Khajeh;Nassoury, Nasha;Tazi, Samia;Giannopoulos, Paresa N.;Guertin, Claude
    • BMB Reports
    • /
    • v.36 no.5
    • /
    • pp.475-487
    • /
    • 2003
  • A gene that encodes a homologue to baculoviral p74, an envelope-associated viral structural protein, has been identified and sequenced on the genome of Choristoneura fumiferana granulovirus (ChfuGV). A part of the ChfuGV p74 gene was located on an 8.9 kb BamHI subgenomic fragment using different sets of degenerated primers. These were designed using the results of the protein sequencing of a major 74 kDa structural protein that is associated with the occlusion-derived virus (ODV). The gene has a 1992 nucleotide (nt) open-reading frame (ORF) that encodes a protein with 663 amino acids with a predicted molecular mass of 74,812 Da. Comparative studies revealed the presence of two major conserved regions in the ChfuGV p74 protein. This study also shows that all of the p74 proteins contain two putative transmembrane domains at their C-terminal segments. At the nucleotide sequence level, two late promoter motifs (TAAG and GTAAG) were located upstream of the first ATG of the p74 gene. The gene contained a canonical poly(A) signal, AATAAA, at its 3' non-translated region. A phylogenetic tree for baculoviral p74 was constructed using a maximum parsimony analysis. The phylogenetic estimation demonstrated that ChfuGV p74 is related the closest to those of Cydia pomonella granulovirus (CpGV) and Phthorimaea operculella granulovirus (PhopGV).

Functional Characterization and Application of the HpOCH2 Gene, Encoding an Initiating $\alpha$l,6-Mannosyltransferase, for N-glycan Engineering in the Methylotrophic Yeast Hansenula polymorpha

  • Kim, Moo-Woong;Kim, Eun-Jung;Kim, Jeong-Yoon;Rhee, Sang-Ki;Kang, Hyun-Ah
    • Proceedings of the Korean Society for Applied Microbiology Conference
    • /
    • 2004.06a
    • /
    • pp.278-281
    • /
    • 2004
  • The $\alpha$1,6-mannosyltransferase encoded by Saccharomyces cerevisiae OCH1 plays a key role for the outer chain initiation of the N-linked oligosaccharides. A search for Hansenula polymorpha genes homologous to S. cerevisiae OCHI (ScOCH1) has revealed seven open reading frames (ORF100, ORF142, ORF168, ORF288, ORF379, ORF576, ORF580). All of the seven ORFs are predicted to be a type II integral membrane protein containing a transmembrane domain near the amino-terminal region and has a DXD motif, which has been found in the active site of many glycosyltransferases. Among this seven-membered OCH1 gene family of H. polymorpha, we have carried out a functional analysis of H. polymorpha ORF168 (HpOCH2) showing the highest identity to ScOCH1. Inactivation of this protein by disruption of corresponding gene resulted in several phenotypes suggestive of cell wall defects, including hypersensitivity to hygromycin B and sodium deoxycholate. The structural analysis of N-glycans synthesized in HpOCH2-disrupted strain (Hpoch2Δ) and the in vitro $\alpha$1,6-mannosyltransferase activity assay strongly indicate that HpOch2p is a key enzyme adding the first $\alpha$1,6-mannose residue on the core glycan Man$_{8}$GlcNAc$_2$. The Hpoch2Δ was further genetically engineered to synthesize a recombinant glycoprotein with the human compatible N-linked oligosaccharide, Man$_{5}$GlcNAc$_2$, by overexpression of the Aspergillus saitoi $\alpha$1,2-mannosidase with the 'HDEL” ER retention signal.gnal.

  • PDF