• Title/Summary/Keyword: Poly-Peptide

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Comparable Electron Capture Efficiencies for Various Protonated Sites on the 3rd Generation Poly(Propylene Imine) Dendrimer Ions: Applications by SORI-CAD and Electron Capture Dissociation Mass Spectrometry (ECD MS)

  • Han, Sang-Yun;Lee, Sun-Young;Oh, Han-Bin
    • Bulletin of the Korean Chemical Society
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    • v.26 no.5
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    • pp.740-746
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    • 2005
  • In this article, we report the tandem mass spectrometry investigations for the electron capture efficiencies of the protons belonging to the different locations (generations) in a poly(propylene imine) dendrimer with three layers of a repeat unit (named as the third generation dendrimer). The employed tandem mass spectrometry methods include SORI-CAD (sustained off-resonance irradiation collisional activation dissociation) and ECD(electron capture dissociation) mass spectrometry. We obtained SORI-CAD spectra for the dendrimer ions in the different charge states, ranging from 2+ to 4+. The analysis of fragmentation sites provides the information as to where the protons are distributed among various generations of the dendrimer. Based upon this, a new strategy to study the electron capture efficiencies of the protons is utilized to examine a new type of triplycharged ions by SORI-CAD, i.e., the 3+ ions generated from the charge reduction of the native 4+ ions by ECD: (M+4H)$^{4+}\;+\;e^-\;{\rightarrow}$ (M+4H)$^{3+\bullet}$ ${\rightarrow}\;({H^{\bullet}}_{ejected}$) + (M+3H)$^{3+}\;\rightarrow$ CAD. Interestingly, comparison of these four SORICAD spectra indicates that the proton distribution in the charge-reduced 3+ ions is very close to that in the native 4+ ions. It further suggests that in this synthetic polymer ($\sim$1.7 kDa) with an artificial architecture, the electron capture efficiencies of the protons are actually insensitive to where they are located in the molecule. This is somewhat contradictory to common expectations that the protons in the inner generations may not be well exposed to the incoming electron irradiation as much as the outer ones are, thus may be less efficient for electron capture. This finding may carry some implications for the case of medium sized peptide ions with similar masses, which are known to show no obvious site-specific fragmentations in ECD MS.

Fungal Growth and Manganese Peroxidase Production in a Deep Tray Solid-State Bioreactor, and In Vitro Decolorization of Poly R-478 by MnP

  • Zhao, Xinshan;Huang, Xianjun;Yao, Juntao;Zhou, Yue;Jia, Rong
    • Journal of Microbiology and Biotechnology
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    • v.25 no.6
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    • pp.803-813
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    • 2015
  • The growth of Irpex lacteus F17 and manganese peroxidase (MnP) production in a selfdesigned tray bioreactor, operating in solid-state conditions at a laboratory scale, were studied. The bioreactor was divided into three layers by three perforated trays. Agroindustrial residues were used both as the carrier of bound mycelia and as a nutrient medium for the growth of I. lacteus F17. The maximum biomass production in the bioreactor was detected at 60 h of fermentation, which was consistent with the CO2 releasing rate by the fungus. During the stationary phase of fungal growth, the maximum MnP activity was observed, reaching 950 U/l at 84 h. Scanning electron microscopy images clearly showed the growth situation of mycelia on the support matrix. Furthermore, the MnP produced by I. lacteus F17 in the bioreactor was isolated and purified, and the internal peptide sequences were also identified with mass spectrometry. The optimal activity of the enzyme was detected at pH 7 and 25℃, with a long half-life time of 9 days. In addition, the MnP exhibited significant stability within a broad pH range of 4-7 and at temperature up to 55℃. Besides this, the MnP showed the ability to decolorize the polymeric model dye Poly R-478 in vitro.

A comprehensive review of techniques for biofunctionalization of titanium

  • Hanawa, Takao
    • Journal of Periodontal and Implant Science
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    • v.41 no.6
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    • pp.263-272
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    • 2011
  • A number of surface modification techniques using immobilization of biofunctional molecules of Titanium (Ti) for dental implants as well as surface properties of Ti and Ti alloys have been developed. The method using passive surface oxide film on titanium takes advantage of the fact that the surface film on Ti consists mainly of amorphous or low-crystalline and nonstoichiometric $TiO_2$. In another method, the reconstruction of passive films, calcium phosphate naturally forms on Ti and its alloys, which is characteristic of Ti. A third method uses the surface active hydroxyl group. The oxide surface immediately reacts with water molecules and hydroxyl groups are formed. The hydroxyl groups dissociate in aqueous solutions and show acidic and basic properties. Several additional methods are also possible, including surface modification techniques, immobilization of poly(ethylene glycol), and immobilization of biomolecules such as bone morphogenetic protein, peptide, collagen, hydrogel, and gelatin.

A feruloyl esterase derived from a leachate metagenome library

  • Rashamuse, Konanani;Sanyika, Walter;Ronneburg, Tina;Brady, Dean
    • BMB Reports
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    • v.45 no.1
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    • pp.14-19
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    • 2012
  • A feruloyl esterase encoding gene (designated fae6), derived from a leachate metagenomic library, was cloned and the nucleotide sequence of the insert DNA determined. Translational analysis revealed that fae6 consists of a 515 amino acid poly-peptide, encoding a 55 kDa pre-protein. The Fae6 primary structure contained the G-E-S-A-G sequence, which corresponds well with a typical catalytic serine sequence motif (G-x-S-x-G). The fae6 gene was successfully over-expressed in E. coli and the recombinant protein was purified to 8.4 fold enrichment with 17% recovery. The $K_M$ data showed Fae6 has a high affinity to methyl sinapate while thermostability data indicated that fae6 was thermolabile with a half life ($T_{1/2}$) < 30 min at $50^{\circ}C$. High affinity for Fae6 against methyl sinapate, methyl ferulate and ethyl ferulate suggest that the enzyme can be useful in hydrolyzing ferulated polysaccharides in a biorefinery process.

Studies on Flacherie and Ina-flacherie Viruses of the Silkworm, Bambyx mori II. Some Properties of Polypeptide of Flacherie Virus (가잠의 연화병 바이러스에 관한 연구 II. 연화병 바이러스 Polypeptide의 성상)

  • 강석권;김근영
    • Journal of Sericultural and Entomological Science
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    • v.21 no.2
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    • pp.7-10
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    • 1979
  • Purified preparations of flacherie virus capsid protein were fractionated by SDS-polyacrylamide gel electrophoresis, and amino acid composition was determined by amino acid analyzer. Three polypeptide components, FP I, FP II and FP III were detected, and the molecular weights of these components were 37,500, 30,500 and 26,500 respectively. The FP III was major poly-peptide comprised about 68.4% of the total virus capsid protein. Seventeen amino acids were detected by an amino acid analyzer from hydrolyzate of the virus capsid protein and the pattern of amino acid composition was similar to those of several other insect viruses.

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Immobilization of Arg-Gly-Asp (RGD) Sequence in Sugar-Containing Copolymer for Culturing Fibroblast Cells

  • Na, Kun;Park, Keun-Hong
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.193-196
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    • 2004
  • The peptide Arg-Gly-Asp (RGD) was immobilized through their amino terminus to ends of a sugar bearing copolymer, producing a functional hybrid copolymer. Poly(N-p-vinylbenzyl-D-maltonamide-co-6-(p-vinylbenzamido)-hexanoic acid-g-GRGDS) [p(VMA-co-VBGRGDS)] promoted the attachment and growth of NIH fibroblast cells. The interaction between fibroblast cells and p(VMA-co- VBGRGDS) copolymer resulted in effective cell attachment, proliferation, and morphological changes by introduction of a GRGDS sequence. Moreover, when pretreated with soluble RGD monomer, attachment of fibroblast cells was suppressed approximately 50% from that of the p(VMA-co-VBGRGDS) surface.

Molecular Cloning, Characterization and Expression Analysis of an ILF2 Homologue from Tetraodon nigroviridis

  • Wang, Hui-Ju;Shao, Jian-Zhong;Xiang, Li-Xin;Shen, Jia
    • BMB Reports
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    • v.39 no.6
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    • pp.686-695
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    • 2006
  • Interleukin-2 enhancer binding factor 2 (ILF2) was reported to regulate transcription of interleukin-2 (IL-2), a central cytokine in the regulation of T-cell responses. This property of ILF2 was well characterized in human and mammals, but little is known in bony fish. In this paper, an ILF2 homologue was cloned and well characterized from Tetraodon nigrovirid is for the further investigation of the function of ILF2 in bony fish. The full-length Tetraodon ILF2 cDNA was 1380 bp in size and contained an open reading frame (ORF) of 1164 bp that translates into a 387 amino-acid peptide with a molecular weight of 42.9 kDa, a 5' untranslated region (UTR) of 57 bp, and a 3' UTR of 159 bp containing a poly A tail. The deduced peptide of Tetraodon ILF2 shared an overall identity of 58%~93% with other known ILF2 sequences, and contained two N-glycosylation sites, two N-myristoylation sites, one RGD cell attachment sequence, six protein kinase C phosphorylation sites, one amino-terminal RGG-rich single-stranded RNA-binding domain, and a DZF zinc-finger nucleic acid binding domain, most of which were highly conserved through species compared. Constitutive expression of Tetraodon ILF2 was observed in all tissues examined, including gill, gut, head kidney, spleen, liver, brain and heart. The highest expression was detected in heart, followed by liver, head kidney and brain. Stimulation with LPS did not significantly alter the expression of Tetraodon ILF2. Gene organization analysis showed that the Tetraodon ILF2 gene have fifteen exons, one more than other known ILF2 genes in human and mouse. Genes up- and down-stream from the Tetraodon ILF2 were Rpa12, Peroxin-11b, Smad4, Snapap and Txnip homologue, which were different from that in human and mouse.

The penetration enhancement and the lipolystic effects of TAT-GKH, in both In vitro, Ex vivo, and In vivo.

  • Lim, J.M.;Chang, M.Y.;Park, S.G.;Kang, N.G.;Song, Y.S.;Lee, Y.H.;Yoo, Y.C.;Cho, W.G.;Han, S.G.;Kang, S.H.
    • Proceedings of the SCSK Conference
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    • 2003.09b
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    • pp.87-107
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    • 2003
  • It was demonstrated that Transactivating transcriptional activator(TAT) protein from HIV-1 shown to enter cells when added to the surrounding media. TAT peptide chemically attached to various proteins was able to deliver these proteins to various cell and even in tissues in mice with high levels in heart and spleen. In this study, the tripeptide GKH(Glycine-Lysine-Histidine) derived from Parathyroid hormone (PTH), which was known as lipolytic peptide, is attached to 9-poly Lysine(TAT) to be used as a cosmetic ingredient for slimming products. When Glycerol release, expressed as extracellular glycerol concentration, is lipolysis index, TAT-GKH at $10^{-5}$mo1/L induces approximately 41.5% maximal lipolytic effects in epididymal adipocytes isolated from rats, compared with basal lipolysis. Epididymal adipose tissues of male rats is assessed ex vivo by microdialysis. Probes are perfused with Ringer solution in which increasing concentrations of TAT-GKH. The perfusion of TAT-GKH induces lipolytic effect. Penetration study showed that TAT-GKH efficiently elevates 36 times higher penetration into the excised hairless mice skin than GKH. in vivo study showed that TAT-GKH had a better effect upon the relative volume of eye bag after 28 days of application on twenty(+2) healthy female volunteers. It was identified that TAT-GKH increases penetration enhancement and lipolytic effects in both in vitro, ex vivo and in vivo.

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Thermoresponsive Graft Copolymers of Hyaluronic Acid (히알루론산의 온도감응성 그래프트 공중합체)

  • Choi, So-Young;Lee, Jong-Hwi
    • Polymer(Korea)
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    • v.35 no.3
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    • pp.223-227
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    • 2011
  • Stimuli-responsive polymers have been investigated as the materials playing the critical roles in various applications. Thermoresponsive graft copolymers, poly (N-isopropylacrylamide)-g-hyaluronic acid (PNIPAAm-g-HA) and elastin-like peptide-g-hyaluronic acid (ELP-g-HA), were synthesized by coupling carboxylic polymers (PNIPAAm-COOH or ELP) to biocompatible HA through amide linkages. Thermoresponsive behavior was observed in both the copolymers, and the results of turbidity measurement were consistent with the results of rheological examination. Among the two copolymers, the ELP graft copolymer shows less cooperative LCST transition than the PNIPAAm case. As the content of graft chains of PNIPAAm and ELP increases, viscosity increases, and the increase was larger in PNIPAAm case at a graft content. These results shows us that the introduction of grafts provides thermosensitivity to biocompatible HA, whose characteristics can be engineered.

Isolation and Phylogenetic Analysis of Acyl-CoA-binding Protein Gene from Panax ginseng C.A. Meyer (인삼으로부터 Acyl-CoA-binding Protein 유전자의 동정 및 계통적 분석)

  • 인준교;류명현;최광태;최관삼;김세영;양덕춘
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.4
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    • pp.201-204
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    • 2001
  • Acyl-CoA binding proteins (ACBP) are small highly-conserved cytosolic proteins that bind long-chain acyl-CoAs. A cDNA encoding ACBP was identified from cDNA library constructed from hairy root poly $A^{+}$ RNA in expressed sequence tags (EST) analysis. The cDNA clone was 453 bp long and carried an open reading frame of 264 bp (10 kDa). The ginseng ACBP amino acid sequence was compared with other reported plant ACBPs using the CLUSTALW. Ginseng ACBP is 89%, 81%, 80%, and 73% identical with ACBP from castor bean, lilly, Digitalis and Arabidopsis, respectively. However, ginseng ACBP is 5 amino acids smaller than Arabidopsis and rape seed ACBPs. Also there is no any known signal peptide sequence in ginseng ACBP.

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