• Title/Summary/Keyword: alanine scanning

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Characterization of Alanine Scanning Mutants of a Peptide Specifically Binding to $TiO_{2}$ Nanoparticles ($TiO_{2}$ Nanoparticle에 특이적으로 결합하는 Peptide의 Alanine Scanning Mutant의 성질에 관한 연구)

  • Seo, Min-Hee;Chael, Hee-Kwon;Myung, Heejoon
    • Microbiology and Biotechnology Letters
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    • v.33 no.4
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    • pp.319-321
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    • 2005
  • We have previously reported the isolation and characterization of peptides binding to $TiO_{2}$ nanoparticles from phage display peptide libraries. One of the peptides (PEP9) was selected and mutant peptide-displaying phages were produced by alanine scanning mutagenesis. The mutant phages were subjected to binding analysis to $TiO_{2}$ nanoparticles. When the proline at residue 4 was substituted by alanine, the binding activity was reduced to $10\%$ of that of wild type PEP9. Substitution of valine at residue 2, serine at residue 3, and isoleucine at residue 5 also decreased the binding to $40\%$. Based on these observations, we concluded that the three dimensional structure generated by residues 2-5 was the critical factor for the binding between PEP9 and the nanoparticle.

Substrate Specificity of Protein Kinase UL97, an antiviral target, on Mutant Peptide Substrates Derived from a Peptide, KESYSVYVYKV (KESYSVYVYKV로부터 변형된 펩타이드 기질을 이용한 항바이러스제의 타깃이 되는 UL97 단백질 인산화 효소의 기질 특이성)

  • Baek, Moon-Chang
    • YAKHAK HOEJI
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    • v.52 no.6
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    • pp.466-470
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    • 2008
  • Human cytomegalovirus expresses an unusual protein kinase UL97, a member of ${H_V}{U_L}$ family of protein kinase. UL97 can phosphorylate nucleoside analogs such as ganciclovir as well as protein/peptide. It has previously been reported that UL97 is able to phosphorylate a KESYSVYVYKV peptide and that P+5 position (K) is important. We examined the extent of contribution of other positions (P-4 through P+6) of the peptide to be substrate of UL97 using alanine substituted peptides (Ala scanning) and deleted peptides. The result suggested that the E (P-2) is negative effect and P+5 (K) is still important. The peptide YSVYVYK is the shortest substrate enough to show high activity, which could be a starting point to develop peptidomimetic drug. This study would give important information to deeply understand the substrate specificity of UL97 and develop an antiviral drug using the small peptide identified here.

PEGylation of Silk Fibroin Model Peptide

  • Kweon, Hae-Yong;Jo, You-Young;Yeo, Joo-Hong;Woo, Soon-Ok;Han, Sang-Mi;Lee, Kwang-Gill
    • International Journal of Industrial Entomology and Biomaterials
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    • v.20 no.2
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    • pp.87-91
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    • 2010
  • Silk fibroin model peptide, alanine pentamer was synthesized through solid-phase method and modified with poly(ethylene glycol). Nuclear magnetic resonance spectrometry and Fourier-transform infrared spectroscopy showed the conformation of alanine pentamer, $\beta$-sheet structure and random coil conformation were not changed with PEGylation. Differential scanning calorimetry showed that relatively strong exothermic peak around $180^{\circ}C$ by PEGylation. No cytotoxicity of PEGylated pentamer was observed by L929 cell proliferation test.

Self-assembly and Mechanism of L-Alanine-based Dihydrazide Derivative as Excellent Gelator of Organic Solvents

  • Wang, Chuan-Sheng;Wang, Xiao-Hong;Li, Zhi-Yuan;Wei, Wei;Shi, Zhong-Liang;Sui, Zhi-Tong
    • Bulletin of the Korean Chemical Society
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    • v.32 no.4
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    • pp.1258-1262
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    • 2011
  • A new organogelator, L-Alanine dihydrazide derivative can self-assemble in various organic solvents and turned them into thermally reversible physical supramolecular organogels at extremely low concentrations (< 2 wt %). The gel-sol phase transition temperatures ($T_{GS}$) were determined as a function of gelator concentration and the corresponding enthalpies (${\Delta}H_g$) were extracted. Scanning electron microscopy (SEM) measurements revealed that the interspaces of fiber-like network structures were diminished with the increasing of the LMOG concentration. FT-IR spectroscopy studies revealed that hydrogen-bonding and hydrophobic interaction were the driving forces for the formation of the gels. Based on the data of XRD and molecular modeling, the possible packing modes for the formation of organogelator aggregates were proposed.

Solution structure and functional analysis of HelaTx1: the first toxin member of the κ-KTx5 subfamily

  • Park, Bong Gyu;Peigneur, Steve;Esaki, Nao;Yamaguchi, Yoko;Ryu, Jae Ha;Tytgat, Jan;Kim, Jae Il;Sato, Kazuki
    • BMB Reports
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    • v.53 no.5
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    • pp.260-265
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    • 2020
  • Scorpion venom comprises a cocktail of toxins that have proven to be useful molecular tools for studying the pharmacological properties of membrane ion channels. HelaTx1, a short peptide neurotoxin isolated recently from the venom of the scorpion Heterometrus laoticus, is a 25 amino acid peptide with two disulfide bonds that shares low sequence homology with other scorpion toxins. HelaTx1 effectively decreases the amplitude of the K+ currents of voltage-gated Kv1.1 and Kv1.6 channels expressed in Xenopus oocytes, and was identified as the first toxin member of the κ-KTx5 subfamily, based on a sequence comparison and phylogenetic analysis. In the present study, we report the NMR solution structure of HelaTx1, and the major interaction points for its binding to voltage-gated Kv1.1 channels. The NMR results indicate that HelaTx1 adopts a helix-loop-helix fold linked by two disulfide bonds without any β-sheets, resembling the molecular folding of other cysteine-stabilized helix-loop-helix (Cs α/α) scorpion toxins such as κ-hefutoxin, HeTx, and OmTx, as well as conotoxin pl14a. A series of alanine-scanning analogs revealed a broad surface on the toxin molecule largely comprising positively-charged residues that is crucial for interaction with voltage-gated Kv1.1 channels. Interestingly, the functional dyad, a key molecular determinant for activity against voltage-gated potassium channels in other toxins, is not present in HelaTx1.

Fabrication of Amino Acid Based Silver Nanocomposite Hydrogels from PVA- Poly(Acrylamide-co-Acryloyl phenylalanine) and Their Antimicrobial Studies

  • Cha, Hyeong-Rae;Babu, V. Ramesh;Rao, K.S.V. Krishna;Kim, Yong-Hyun;Mei, Surong;Joo, Woo-Hong;Lee, Yong-Ill
    • Bulletin of the Korean Chemical Society
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    • v.33 no.10
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    • pp.3191-3195
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    • 2012
  • New silver nanoparticle (AgNP)-loaded amino acid based hydrogels were synthesized successfully from poly (vinyl alcohol) (PVA) and poly(acryl amide-co-acryloyl phenyl alanine) (PAA) by redox polymerization. The formation of AgNP in hydrogels was confirmed by using a UV-Vis spectrophotometer and XRD. The structure and morphology of silver nanocomposite hydrogels were studied by using a scanning electron microscopy (SEM), which demonstrated scattered nanoparticles, ca. 10-20 nm. Thermogravimetric analysis revealed large differences of weight loss (i.e., 48%) between the prestine hydrogel and silver nanocomposite. The antibacterial studies of AgNP-loaded PAA (Ag-PAA) hydrogels was evaluated against Escherichia coli (Gram-negative) and Staphylococcus aureus (Gram-positive) bacteria. These Ag-PAA hydrogels showed significant activities against all the test bacteria. Newly developed hydrogels could be used for medical applications, such as artificial burn dressings.

Optimization of Scan Parameters for in vivo Hyperpolarized Carbon-13 Magnetic Resonance Spectroscopic Imaging

  • Nguyen, Nguyen Trong;Rasanjala, Onila N.M.D.;Park, Ilwoo
    • Investigative Magnetic Resonance Imaging
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    • v.26 no.2
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    • pp.125-134
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    • 2022
  • Purpose: The aim of this study was to investigate the change in signal sensitivity over different acquisition start times and optimize the scanning window to provide the maximal signal sensitivity of [1-13C]pyruvate and its metabolic products, lactate and alanine, using spatially localized hyperpolarized 3D 13C magnetic resonance spectroscopic imaging (MRSI). Materials and Methods: We acquired 3D 13C MRSI data from the brain (n = 3), kidney (n = 3), and liver (n = 3) of rats using a 3T clinical scanner and a custom RF coil after the injection of hyperpolarized [1-13C]pyruvate. For each organ, we obtained three consecutive 3D 13C MRSI datasets with different acquisition start times per animal from a total of three animals. The mean signal-to-noise ratios (SNRs) of pyruvate, lactate, and alanine were calculated and compared between different acquisition start times. Based on the SNRs of lactate and alanine, we identified the optimal acquisition start timing for each organ. Results: For the brain, the acquisition start time of 18 s provided the highest mean SNR of lactate. At 18 s, however, the lactate signal predominantly originated from not the brain, but the blood vessels; therefore, the acquisition start time of 22 s was recommended for 3D 13C MRSI of the rat brain. For the kidney, all three metabolites demonstrated the highest mean SNR at the acquisition start time of 32 s. Similarly, the acquisition start time of 22 s provided the highest SNRs for all three metabolites in the liver. Conclusion: In this study, the acquisition start timing was optimized in an attempt to maximize metabolic signals in hyperpolarized 3D 13C MRSI examination with [1-13C] pyruvate as a substrate. We investigated the changes in metabolic signal sensitivity in the brain, kidney, and liver of rats to establish the optimal acquisition start time for each organ. We expect the results from this study to be of help in future studies.

Serum Biochemical, Histopathology and SEM Analyses of the Effects of the Indian Traditional Herb Wattakaka Volubilis Leaf Extract on Wistar Male Rats

  • Gopal, Velmani;Mandal, Vivekananda;Tangjang, Sumpam;Mandal, Subhash C.
    • Journal of Pharmacopuncture
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    • v.17 no.1
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    • pp.13-19
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    • 2014
  • Objectives: The present study investigated the protective effect of Wattakaka (W.) volubilis leaf extract against streptozotocin (STZ)-induced diabetes in rats. Methods: Male Wistar rats were divided into five groups (with six rats in each group) and were fed ad libitum. The rats were fasted for sixteen hours before diabetes was induced by injecting a single dose of 90 mg/kg body weight of STZ in 0.9-percent normal saline through an intraperitoneal route. The five groups were as follows: Group 1: normal control (saline-treated), Group 2: untreated diabetic rats, Groups 3 and 4: diabetic rats treated orally with petroleum ether cold maceration extract (PEME) of W. volubilis (50 and 100 mg/kg body weight), and Group 5: diabetic rats treated orally with metformin (250 mg/kg body weight). All rats received treatment for 21 days. For the STZ-induced diabetic rats, the blood-glucose, ${\alpha}$-amylase, total protein and alanine transaminase (ALT) levels were measured on days 7, 14 and 21 of the treatment with PEME of W. volubilis and the treatment with metformin. Histopathological changes in the liver were examined with hematoxylin-eosin staining. Morphological changes in the liver were also examined with glutaraldehyde fixation. Results: The treatments with PEME of W. volubilis and with metformin in experimental rats by oral injections for 21 days produced reductions in the levels of serum biochemical markers. Histopathology and scanning electron microscopy results showed that the administrations of PEME of W. volubilis and of metformin suppressed the generation of abnormal liver cells in the STZ-treated rats. Conclusion: These results suggest that both PEME of W. volubilis and metformin have a protective effect against STZ-induced diabetes.

Effect of High Dietary Copper on the Morphology of Gastro-Intestinal Tract in Broiler Chickens

  • Chiou, P.W.S.;Chen, C.L.;Chen, K.L.;Wu, C.P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.4
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    • pp.548-553
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    • 1999
  • An experiment was conducted to study the effects of high dietary copper supplementation on the gastrointestinal tract morphology of broiler chickens. Eighty 3-week-old broiler chicks were divided randomly into eight groups of four dietary treatments and over three week were fed isoenergetic and isonitrogenous diets that contained 0, 100, 250, or 500 mg/kg of supplemental copper from cupric sulfate. The copper supplementation in the broiler diet up to 250 mg/kg did not significantly influence broilers' performance. A high dietary copper supplementation of 500 mg/kg did significantly depress growth and feed conversion in the broilers (p<0.05). Copper supplementation more than 250 mg/kg in the broiler diet significantly influenced the morphology of the GI tract, as shown by severe oral lesions and gizzard erosion. It also significantly depressed the villi height and significantly thickened the muscular layer in the duodenum (p<0.05). The severely damaged villi were observed by scanning electronic microscope from the duodenum samples of broilers fed a 500 mg/kg copper supplemented diet. The 500 mg/kg copper supplemented diet also significantly influenced the plasma constituents. Plasma glucose concentration was significantly depressed (p<0.05).

Depigmentation Activity of Barley, Unpolished Rice, Job's-tear (보리·현미·율무의 미백효과에 관한 연구)

  • Lee, Tae-huen;Kim, Hye-jeong;Kim, Yoon-bum
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.16 no.2
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    • pp.57-77
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    • 2003
  • Objectives: The aim of this study was to investigate the depigmentation effects of Barley, Unpolished rice, Job's-tear. Metbods: We investigated that the extracts of Barley, Unpolished rice, Job's-tear inhibit activity of tyrosinase, the enzyme which convert ts 3-(3,4-dihydroxyphenyl) alanine to dopachrom in the biosynthetic process of melanin, the UV absorbance of the extracts in the UV-A region and UV-B region was measured by UV scanning, the effect of extracts on cell viability and melanin production in cultured B16 mouse melanoma cells were measured, and cytoprotective effects of extracts on PC12 cells injured by hydrogen peroxide measured by MTT assay. Results: The extracts of Barley, Unpolished rice, Job's-tear inhibited activity of tyrosinase in low density. The Barley, Job's-tear extracts not only showed inhibitory effects on melanin production in cultured B16 mouse melanoma cells, but also exhibited cytoprotective effects on PC12 cells injured by hydrogen peroxide in low density. Unpolished rice extract showed inhibitory effect on melanin production in cultured B16 mouse melanoma cells, but did not showed cytoprotective effect Barley, Unpolished rice, Job's-tear extracts did not showed an absorbance effect in the UV-A region and UV-8 region. Conclusions: These results suggest that Barley, Unpolished rice, Job's-tear inhibit melanin biosynthesis which is involved in hyperpigmentation and could be used as a whitening agent.

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