• Title/Summary/Keyword: a synthetic peptide

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The C-terminal Region of Human Tau Protein with Ability of Filament Formation

  • Chung, Sang-Ho
    • Animal cells and systems
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    • v.1 no.2
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    • pp.317-321
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    • 1997
  • Tau protein is one of the microtubule-associated proteins in the mammalian brain. In Alzheimer's disease, tau protein is immobilized in the somatodendritic compartment of certain nerve cells, where it forms a part of the paired helical filament (PHF). To understand the role of tau protein in the formation of PHF, a recombinant human tau protein expressed in Escherichia coli and five synthetic peptide fragments (peptide 1 to peptide 5), corresponding to the C-terminal region of tau protein, were prepared and their ability in self-assembly to form filamentous structures was examined. The recombinant human tau protein formed short rod-like structures in 0.1M MES buffer containing 1 mM $MgCI_2$, while a synthetic peptide fragment 1 containing 55 amino acid residues could assemble into a lot of long filamentous structures in water and particularly twisted helical structures in 0.1M MES buffer containing 1 mM $MgCI_2$. This suggests that the C-terminal region possesses a filament-forming ability and may be related to the formation of the helical structure by providing a powerful filament-forming driving force.

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Immunohistochemical Detection of N-myc Gene Product by Using Antiserum Against Synthetic Peptide (항-펩타이드 항체를 이용한 암유전자 N-myc 산물의 면역조직화학적 검출)

  • Lee, Hyun-Chul;Lee, Wan-Joo;Ahn, Tai-Hew
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.2
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    • pp.167-174
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    • 1987
  • N-myc, a DNA sequence related to the oncogene c-myc, was found to be amplified in untreated primary neuroblastomas and the amplification appeared to be associated with advanced disease at diagnosis and rapid tumor progression. Synthetic peptides have been useful immunogens for generating antisera and monoclonal antibodies to a number of native proteins. In order to identify myc-related protein in the tumor cells, an antiserum against a synthetic hexapeptide (-Glu-Asp-Ile-Trp-Lys-Lys-), whose sequence corresponds to a part of the exon 2 of oncogene N-myc, was prepared by immunizing a rabbit with BSA-conjugated peptide. After ammonium sulfate precipitation and affinity column chromatography, it appeared to be specific to the peptide. Strong nuclear staining in immunoperoxidase method using this serum was observed in both human promyeloid leukemic cell line, HL-60(containing high c-myc copy number), and human neuroblastoma cell line, LA-N-5 (containing high N-myc copy number), whereas LA351 (human lymphoid cell line) cells did not react with the serum. This reaction was completely abrogated by incubating the antiserum with soluble excess peptide. These data suggest that the protein encoded by N-myc could be localized in the nucleus as c-myc protein and this antiserum can be used to detect myc-related tumor cells in clinical samples and to determine if the N-myc expression correlates with genomic amplification in cell lines, untreated primary tumors, and untreated metastases.

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The High Production of Multimeric Angiotensin-converting-enzyme-inhibitor in E. coli

  • Park Je-Hyoen;Kim Sun-Hoi;Ahn Sun-Hee;Lee Jong-Hee;Kim Young-Sook;Lee Sang-Jun;Kong In-Soo
    • Fisheries and Aquatic Sciences
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    • v.4 no.2
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    • pp.84-87
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    • 2001
  • Multimeric angiotensin-converting-enzyme-inhibitor (ACE}) containing a trypsin cleavable linker peptide between ACEI was constructed. We made synthetic DNA coding for the ACEI peptide with asymmetric and complementary cohesive ends of linker nucleotides. A tandemly repeated DNA cassette for the expression of concatameric short peptide multimers was constructed by ligating the basic units. The resultant multimeric peptide expressed as soluble and trypsin treated peptide was shown at the same retention time with chemically synthetic ACEI by HPLC. The present results showed that the technique developed for the production of the ACEI multimers with trypsin cleavable linker peptides can be generally applicable to the production of short peptide.

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Synergistic Killing Effect of Synthetic Peptide P20 and Cefotaxime on Methicillin-Resistant Nosocomial Isolates of Staphylococcus aureus

  • Jung, Hyun-Jun;Choi, Kyu-Sik;Lee, Dong-Gun
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1039-1046
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    • 2005
  • The salt resistance of antibacterial activity and synergistic effect with clinically used antibiotic agents are critical factors in developing effective peptide antibiotic drugs. For this reason, we investigated the resistance of antibacterial activity to antagonism induced by NaCl and $MgCl_2$ and the synergistic effect of P20 with cefotaxime. P20 is a 20-residue synthetic peptide derived from a cecropin A (CA)-melittin(ME) hybrid peptide. In this study, P20 was found to have potent antibacterial activity against clinically isolated methicillin-resistant Staphylococcus aureus (MRSA) strains without hemolytic activity against human erythrocytes. The combination study revealed that P20 in combination with cefotaxime showed synergistic antibacterial activity in an energy-dependent manner. We also confirmed the synergism between P20 and cefotaxime by fluorescence-activated flow cytometric analysis by staining bacterial cells with propidium iodide (PI) and bis-(1,3-dibutylbarbituric acid) trimethine oxonol (BOX). This study suggests that P20 may be useful as a therapeutic antibiotic peptide with synergistic effect in combination with conventional antibiotic agents.

Determination of the minimal sequence of bovine lactoferricin responsible for apoptosis induction

  • Yoo, Yung-Choon;Lee, Kyung-Bok;Lee, Hoi-Young
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.134.2-135
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    • 2003
  • We examined the minimal amino acid sequence of bovine lactoferricin (Lfcin-B), a cationic peptide corresponding to residues 17-41 near the N-terminus of bovine lactoferrin, to induce apoptosis in THP-l human monocytic leukemic cells using synthetic peptides. A synthetic peptide (Lfc-17/29, amino acid sequence; FKCRRWQWRMKKL) which is consist of 13 amino acids near the N-terminus of Lfcin-B induced cell death in THP-1 cells in a dose-dependent manner, showing apparent apoptotic changes such as hypodiploid forms of genomic DNA and apoptotic DNA fragmentation. (omitted)

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Inhibitory Effects of Synthetic Peptides Containing Bovine Lactoferrin C-lobe Sequence on Bacterial Growth

  • Kim, Woan-Sub;Ohashi, Midori;Shimazaki, Kei-ichi
    • Food Science of Animal Resources
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    • v.36 no.4
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    • pp.452-457
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    • 2016
  • Lactoferrin is a glycoprotein with various biological effects, with antibacterial activity being one of the first effects reported. This glycoprotein suppresses bacterial growth through bacteriostatic or bactericidal action. It also stimulates the growth of certain kinds of bacteria such as lactic acid bacteria and bifidobacteria. In this study, Asn-Leu-Asn-Arg was selected and chemically synthesized based on the partial sequences of bovine lactoferrin tryptic fragments. Synthetic Asn-Leu-Asn-Arg suppressed the growth of Pseudomonas fluorescens, P. syringae and Escherichia coli. P. fluorescens is a major psychrotrophic bacteria found in raw and pasteurized milk, which decreases milk quality. P. syringae is a harmful infectious bacterium that damages plants. However, synthetic Asn-Leu-Asn-Arg did not inhibit the growth of Lactobacillus acidophilus. It is expected that this synthetic peptide would be the first peptide sequence from the bovine lactoferrin C-lobe that shows antibacterial activity.

Identification of a Protein Kinase using a FITC-labelled Synthetic Peptide in Streptomyces griseus IFO 13350 (형광 Peptide를 이용한 Streptomyces griseus IFO 13350의 인산화 단백질 동정)

  • 허진행;정용훈;김종희;신수경;현창구;홍순광
    • Microbiology and Biotechnology Letters
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    • v.30 no.3
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    • pp.235-240
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    • 2002
  • Streptomycetes is a group of Gram-positive soil bacteria that growas a branching vegetative mycelium leading to the formation of spores, and display a physiological differenti-ation related to the synthesis of many secondary metabolites including antibiotics. Their complex life cycle and multicellular differentiation require various levels of regulation and types of signal transduction systems including eukaryotic-type serine/threonine protein kinases and prokaryotic-type histidine/aspartic acid protein kinases. Akt kinase that was found in cells is a sorine/threonine kinase controlling signal pathway for multi-tude of important cellular events. The activation or inactivation of Akt kinase in the cell is one of the critical regulatory points to deliver cell proliferation, differentiation, survival or apoptosis signal. To find the regula-tory protein homologous to Akt in Streptomyces, the fluorescien-labeled synthetic peptide (FITC-TRRSR-TESIT) was designed from the consensus sequence of target proteins for Akt kinase. From the difference of the mobility between the nonphosphorylated and phosphorylated synthetic peptides on Agarose gel electro-phoresis, the Akt-phosphorylating activity was monitored. The cell-free extract prepared from Streptomyces griseus IFO 13350 and the Akt homologous protein was purified by ammonium sulfate fractionation and many steps of column chromatographies such as, DEAE-Sepharose, Mono Q, Resource Phenyl-Soporose and Gel permeation column chromatographies. As a result, the protein phosphorylating the fluorescien-labeled Akt substrate was identified and it's molecular weight was estimated as 39 kDa on SDS-PAGE.

Improvement of osteogenic potential of biphasic calcium phosphate bone substitute coated with synthetic cell binding peptide sequences

  • Choi, Hyunmin;Park, Nho-Jae;Jamiyandorj, Otgonbold;Hong, Min-Ho;Oh, Seunghan;Park, Young-Bum;Kim, Sungtae
    • Journal of Periodontal and Implant Science
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    • v.42 no.5
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    • pp.166-172
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    • 2012
  • Purpose: The aim of this study was to evaluate the improvement of osteogenic potential of biphasic calcium phosphate (BCP) bone substitute coated with synthetic cell-binding peptide sequences in a standardized rabbit sinus model. Methods: Standardized 6-mm diameter defects were created bilaterally on the maxillary sinus of ten male New Zealand white rabbits, receiving BCP bone substitute coated with synthetic cell binding peptide sequences on one side (experimental group) and BCP bone substitute without coating (control group) on the other side. Histologic and histomorphometric analysis of bone formation was carried out after a healing period of 4 or 8 weeks. Results: Histological analysis revealed signs of new bone formation in both experimental groups (4- and 8-week healing groups) with a statistically significant increase in bone formation in the 4-week healing group compared to the control group. However, no statistically significant difference in bone formation was found between the 8-week healing group and the control group. Conclusions: This study found that BCP bone substitute coated with synthetic cell-binding peptide sequences enhanced osteoinductive potential in a standardized rabbit sinus model and its effectiveness was greater in the 4-week healing group than in the 8-week healing group.

Identification of Novel Bioactive Hexapeptides Against Phytopathogenic Bacteria Through Rapid Screening of a Synthetic Combinatorial Library

  • Choi, Jae-Hyuk;Moon, Eun-Pyo
    • Journal of Microbiology and Biotechnology
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    • v.19 no.8
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    • pp.792-802
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    • 2009
  • Antimicrobial peptides (AMPs) are considered to be a promising alternative to conventional antibiotics for future generations. We identified four novel hexapeptides with antimicrobial activity: KCM11 (TWWRWW-$NH_2$), KCM12 (KWRWlW-$NH_2$), KCM21 (KWWWRW-$NH_2$), and KRS22 (WRWFIH-$NH_2$), through positional scanning of a synthetic peptide combinatorial library (PS-SCL). The ability of these peptides to inhibit the growth of a variety of bacteria and unicellular fungi was evaluated. KCM11 and KRS22 preferentially inhibited the normal growth of fungal strains, whereas KCM12 and KCM21 were more active against bacterial strains. Bactericidal activity was addressed in a clear zone assay against phytopathogenic bacteria, including Pectobacterium spp., Xanthomonas spp., Pseudomonas spp., etc. KCM21 showed the highest activity and was effective against a wide range of target organisms. Application of KCM21 with inoculation of Pectobacterium carotovorum subsp. carotovorum on detached cabbage leaves resulted in an immune phenotype or a significant reduction in symptom development, depending on the peptide concentration. Cytotoxicity of the four hexapeptides was evaluated in mouse and human epithelial cell lines using an MTT test. The results revealed a lack of cytotoxic effects.