• Title/Summary/Keyword: amino acid complex

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NMR peak assignment for the elucidation of the solution structure of T4 Endonuclease V

  • Im, Hoo-Kang;Hyungmi Lihm;Yu, Jun-Suk;Lee, Bong-Jin
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.183-183
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    • 1996
  • Bacteriophage T4 endonuclease V initiates the repair of ultraviolet (UV)-induced pyrimidine dimer photoproducts in duplex DNA. The mechanism of DNA strand cleavage involves four sequential stens: linear diffusion along dsDNA, pyrimidine dimer-specific binding,l pyrimidine dimer-DNA glycosylase activity, and Af lyase activity. Although crystal structure is known for this enzyme, solution structure has not been yet known. In order to elucidate the solution structure of this enzyme NMR spectroscopy was used. As a basis for the NMR peak assignment of the protein, HSQC spectrum was obtained on the uniformly $\^$15/N-labeled T4 endonuclease V. Each amide peak of the spectrum were classified according to amino acid spin systems by interpreting the spectrum of $\^$15/N amino acid-specific labeled T4 endonuclease V. The assignment was mainly obtained from three-dimensional NMR spectra such as 3D NOESY-HMQC, 3D TOCSY-HMQC. These experiments were carried out will uniformly $\^$15/N-labeled sample. In order to assign tile resonance of backbon atom, triple-resonance theree-dimensional NMR experiments were also performed using double labeled($\^$15/N$\^$13/C) sample. 3D HNCA, HN(CO)CA, HNCO, HN(CA)HA spectra were recorded for this purpose. The results of assignments were used to interpret the interaction of this enzyme with DNA. HSQC spectrum was obtained for T4 endonuclease V with specific $\^$15/N-labeled amino acids that have been known for important residue in catalysis. By comparing the spectrum of enzyme*DNA complex with that of the enzyme, we could confirm the important role of some residues of Thr, Arg, Tyr in activity. The results of assignments were also used to predict the secondary structure by chemical shift index (CSI).

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Processing and Quality Characteristics of Sea Mussel Mytilus edulis Sauces from Sea Mussel Complex Extract (진주담치(Mytilus edulis) 복합추출물을 이용한 진주담치소스의 제조 및 품질특성)

  • Kim, Seon-Geun;Cho, Jun-Hyun;Hwang, Seok-Min;Nam, Hyeon-Gyu;Choi, Jong-Duck;Oh, Kwang-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.6
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    • pp.656-661
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    • 2017
  • To develop a highly value-added product from extract from small and damaged sea mussels Mytilus edulis, we prepared two types of sea mussel sauce (MS): bottled (BMS) and retort pouched (RMS). We investigated the processing conditions, quality metrics and flavor compounds in each type of sauce. We found that the most appropriate base formulation for both BMS and RMS consisted of 40.0% SME (Brix $30^{\circ}$), 15.0% sugar, 6.0% salt, 4.0% monosodium glutamate, 4.0% soy sauce, 3.5% starch, 3.0% yeast extract, 3.5% wheat flour and 21.0% water. The crude protein, salinity, volatile basic nitrogen and amino-nitrogen content of the BMS and RMS were 8.7% and 8.8%, 9.3% and 9.2%, 24.9 and 31.4 mg/100 g, and 468.5 and 455.1 mg/100 g, respectively. For comparison, the ranges of these values in commercial oyster sauces (COS) are 4.7-7.5%, 10.7-12.0%, 8.2-12.5 mg/100 g, and 225.7-448.2 mg/100 g, respectively. The total free amino acid content of RMS and Premium COS was 7,215.7 and 6,160.7 mg/100 g, respectively, and the main free amino acids were glutamic acid, taurine, glycine, alanine, arginine, proline and lysine. These results demonstrate that BMS and RMS have favorable organoleptic qualities and good storage stability compared to COS, and are suitable for commercialization as high-flavor seasoning sauces.

Analysis of Phosphorylation of the BRI1/BAK1 Complex in Arabidopsis Reveals Amino Acid Residues Critical for Receptor Formation and Activation of BR Signaling

  • Yun, Hye Sup;Bae, Young Hee;Lee, Yun Ji;Chang, Soo Chul;Kim, Seong-Ki;Li, Jianming;Nam, Kyoung Hee
    • Molecules and Cells
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    • v.27 no.2
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    • pp.183-190
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    • 2009
  • The plasma membrane-localized BRASSINOSTEROID-INSENSITIVE1 (BRI1) and BRI1-ASSOCIATED KINASE1 (BAK1) are a well-known receptor pair involved in brassinosteroids (BR) signaling in Arabidposis. The formation of a receptor complex in response to BRs and the subsequent activation of cytoplasmic domain kinase activity share mechanistic characteristics with animal receptor kinases. Here, we demonstrate that BRI1 and BAK1 are BR-dependently phosphorylated, and that phosphorylated forms of the two proteins persist for different lengths of time. Mutations of either protein abolished phosphorylation of the counterpart protein, implying transphosphorylation of the receptor kinases. To investigate the specific amino acids critical for formation of the receptor complex and activation of BAK1 kinase activity, we expressed several versions of BAK1 in yeast and plants. L32E and L46E substitutions resulted in a loss of binding of BAK1 to BRI1, and threonine T455 was essential for the kinase activity of BAK1 in yeast. Transgenic bri1 mutant plants overexpressing BAK1(L46E) displayed reduced apical dominance and seed development. In addition, transgenic wild type plants overexpressing BAK1(T455A) lost the phosphorylation activity normally exhibited in response to BL, leading to semi-dwarfism. These results suggest that BAK1 is a critical component regulating the duration of BR efficacy, even though it cannot directly bind BRs in plants.

Separation of Optical Isomers of DNS-Amino Acids in High-Performance Liquid Chromatography (고성능 액체크로마토 그래피에 의한 Dansyl-아미노산 광학이성질체의 분리)

  • Sun Haing Lee;Tae Sub O;Kyung Sug Park
    • Journal of the Korean Chemical Society
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    • v.30 no.2
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    • pp.216-223
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    • 1986
  • Separation of optical isomers of DNS derivatized amino acids by a reversed-phase high-performance liquid chromatography has been studied by adding a complex of an optically active amino acid (L-arginine) with the metal ion (Cu(II), Zn(II), Cd(II), Ni(II)) to the mobile phase. The separations are affected by the concentrations of acetonitrile, chelate and buffer. They are also affected by the pH and the kinds of metal and buffer. A separation mechanism, which is based on steric effect of the ligand exchange reaction for the formation of ternary complexes by the D,L-DNS-amino acids and the chiral additive associated with the stationary phase, is proposed to interpret the elution behaviors of D, L-dansyl-amino acids.

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Purification and Characterization of the Red Carotenoprotein from the Skin of Ascidian, Halocynthia roretzi 1. Purification and Characterization of the Caritenopritein (멍게껍질로부터 분리제정한 적색 Carotenoprotein의 특성 1. Carotenoprotein 의 정제 및 특성)

  • Kang, Ok-Ju;Suh, Myung-Ja;Lee, An-Jong;Kim, Se-Kwon
    • Journal of Life Science
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    • v.5 no.4
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    • pp.170-180
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    • 1995
  • A carotennnoprotein from the skin of Ascidian(Halocynthia roretzi) was extracted by Triton X-100 and purified by ammonium sulfate fraction, SephadexG-200 charomatography and DEAE-cellulose ion exchange chromatography. The carotenoprotein was redwith broad $\lambda$$_{max}$ between 495, 467 and 318nm. The red carotenoprotein had an approximate molecular weight of 326KDa(gel filtration). SDS-PAGE indicated the presence of two polypeptodes of 84.1KDa and 74.4KDa, with different mobility in polyacrylamide gel electrophoresis. In the presence of denaturing agents such as organic solvent aand extreme pH, the red complex readily disociates to liberate the yellow carotenoid($\lambda$$_{max}$ 452nm) and a colourless apoprotein. The amino acid composition of carotenoprotein were mainly threonine(15.2%), aspartic acid(12.2%), glutamic acid(11.9%) and serine(9.6%), while proline was not found. The carotenoprotein consisted of lipids as structure units. Its major fatty acids composion were C$_{18:1}$, C$_{16:1}$, and C$_{16:0}$. The monounsaturated fatty acids(41.5%) contained abundant content compared to other fatty aacids(polyunsaturated fatty acids 37.4%, saturated fatty acids 20.6%).

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Acrylic Polymer Composition Suitable for Ion Delivery and Selective Detection of Proton, and Hydroxyl and Cu(II) Ions (이온 이동에 적합한 아크릴고분자 박막의 조성과 수소이온, 수산화이온, 구리이온의 선택적 검출)

  • Lee, Dahye;Woo, Heejung;Do, Jung Yun
    • Polymer(Korea)
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    • v.38 no.6
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    • pp.801-808
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    • 2014
  • Ion diffusion and chemical binding to acrylic polymer were investigated in a solid film. The composition of acrylic monomers containing amino group and carboxylic acid was adjusted for rapid ion migration in the film. p-Methylred (PMR) and phenolphthalein derivatives were optically sensitive to the concentration of proton and hydroxyl anion, respectively and verified the ion migration through the film layers. A rapid proton migration was observed in the film of a high amino content. On the other hand, $OH^-$ migration occurred rapidly in a high content of carboxylic acid group. The proton migration occurred through the internal layer as well as surface layer of a film and was reversible during 50 repetition examination. Copper(II) ion migration was examined with a Rhodamine-containing polymer film. The light absorption and emission spectra of a Rhodamine-Cu complex showed the key contribution of carboxylic acid group to the Cu(II) migration in a film.

Electrochemical and Raman Spectroscopy Analysis for D- and L-Tryptophan-b-Cyclodextrin Inclusion Complexes

  • Jeong, Yu-Ra;Lee, So-Ra;Son, Pyeong-Soo;Choi, Seong-Ho
    • Journal of the Korean Applied Science and Technology
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    • v.32 no.3
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    • pp.451-460
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    • 2015
  • An enantioselective recognition of D- and L-tryptophan (Trp)-b-cyclodextrin (CD) inclusion complex was performed using electrochemical and FT-Raman spectroscopic analysis. From the electrochemical analysis, the selectivity coefficient ($K_{DL}$) of b-CD inclusion complexes was found higher than that of the D- and L-Trp in phosphate buffered saline (PBS, pH=7.0) solution. The percentage of enantioselectivity ($I_{%{ee}}$) for peak current of D-Trp-b-CD inclusion complexes was observed higher than that of L-Trp-b-CD inclusion complexes in PBS solution. From Raman spectroscopy, chemical shift difference (D, $cm^{-1}$) for the C=C stretch, ring vibration, and ring breathing of D-Try-b-CD inclusion complex were observed higher than that of L-Trp-b-CD inclusion complex. The electrochemical and Raman spectroscopic analyses were found very useful for chiral detection of racemic amino acid in the presence of b-CD.

Previous Infection with Plasmodium berghei Confers Resistance to Toxoplasma gondii Infection in Mice

  • Lee, Dong-Hun;Chu, Ki-Back;Kang, Hae-Ji;Lee, Su-Hwa;Quan, Fu-Shi
    • Parasites, Hosts and Diseases
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    • v.57 no.2
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    • pp.93-99
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    • 2019
  • Both Plasmodium spp. and Toxoplasma gondii are important apicomplexan parasites, which infect humans worldwide. Genetic analyses have revealed that 33% of amino acid sequences of inner membrane complex from the malaria parasite Plasmodium berghei is similar to that of Toxoplasma gondii. Inner membrane complex is known to be involved in cell invasion and replication. In this study, we investigated the resistance against T. gondii (ME49) infection induced by previously infected P. berghei (ANKA) in mice. Levels of T. gondii-specific IgG, IgG1, IgG2a, and IgG2b antibody responses, $CD4^+$ and $CD8^+$ T cell populations were found higher in the mice infected with P. berghei (ANKA) and challenged with T. gondii (ME49) compared to that in control mice infected with T. gondii alone (ME49). P. berghei (ANKA) + T. gondii (ME49) group showed significantly reduced the number and size of T. gondii (ME49) cysts in the brains of mice, resulting in lower body weight loss compared to ME49 control group. These results indicate that previous exposure to P. berghei (ANKA) induce resistance to subsequent T. gondii (ME49) infection.

The Development of a Natural Seasoning Using the Enzymatic Hydrolysate of Fish Skin (어피의 효소적 가수분해물을 이용한 천연조미료의 개발)

  • 김세권;양현필이응호
    • KSBB Journal
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    • v.6 no.4
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    • pp.327-336
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    • 1991
  • A study on the optimum hydrolysis conditions of fish skin through the aid of enzymes and the development of a natural seasoning using the hydrolysate has been carried out for the effective utilization of fish skin. Using the "pH-drop" techniques the collagenase and pronase were identified as most suitable for this purpose. The $K_m$ and $V_{max}$ values of pronase were 1.82 mgN/ml and 0.06 mgN/mL/min, respectively. The hydrolysis conditions of the cod skin for the pronase were as follows: reaction temperature, $50^{\circ}C$; reaction time, 3hrs; pH 6; enzyme concentration, 0.03%. The degree of hydrolysis at these conditions was 76.8%. But after hydrolyzing cod skin with collagenase for 1hr, when the pronase was treated, the degree of hydrolysis was 83.13%. The molecular weight of the hydrolysate was 8,000 daltons. Among the amino acids in the hydrolysate, glycine(27.95%), glutamic acid(10.94%), proline(7.39%), aspartic acid(9.47%) and serine(7.39%) were responsible for 64.23% of the total amino acids. But valine, methionine, isoleucine, leucine, phenylalanine and histidine having a bitter taste were only 13.05%. From the results of the sensory evaluation, the imitation sauce which was made of 20% fermented soy sauce prepared from the hydrolysate was at least similar to the traditional soybean sauce in product quality. The complex seasoning containing 31.7% of the hydrolysate was nearly equal to complex seasonings on the market, too.

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Heterocyclic Amines Removal by Binding Ability of Lactic Acid Bacteria Isolated from Soybean Paste (된장에서 분리된 유산균의 결합력에 의한 Heterocyclic Amines 제거)

  • Lim, Sung-Mee
    • Korean Journal of Microbiology
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    • v.50 no.1
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    • pp.73-83
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    • 2014
  • The objective of the this study was to investigate the binding capacity and removal ability of lactic acid bacterial strains obtained from Korean soybean paste for mutagenic heterocyclic amines (HCAs) formed during cooking of protein-rich food at high temperature. Among 19 strains identified by carbohydrate fermentation and 16S rRNA sequencing, the live cell or cell-free culture supernatant of Lactobacillus acidophilus D11, Enterococcus faecium D12, Pediococcus acidilactici D19, L. acidophilus D38, Lactobacillus sakei D44, Enterococcus faecalis D66, and Lactobacillus plantarum D70 inhibited the mutagenesis caused by either 3-amino-1,4-dimethyl-5H-pyrido[4,3-b] indole (Trp-P-1) or 3-amino-1-methyl-5H-pyrido[4,3-b] indole (Trp-P-2) in Salmonella typhimurium TA98 and TA100. The bacterial cells of the isolated strains showed greater binding activity than the pure cell wall, exopolysaccharide, and pepetidoglycan. The carbohydrate moieties of the cell wall or protein molecules on the cell surface have a significant role in binding Trp-P-1 and Trp-P-2, since protease, heating, sodium metaperiodate, or acidic pH treatments significantly (P<0.05) reduced the binding efficacy of the tested bacteria. Addition of metal ions or sodium dodecyl sulfate decreased the binding ability of E. faecium D12, L. acidophilus D38, and E. faecalis D66. Therefore, the binding mechanisms of these strains may consist of ion-exchange and hydrophobic bonds. Especially, the high mutagen binding by L. acidophilus D38 and L. plantarum D70 may reduce the accumulation or absorption of Trp-P-1 and Trp-P-2 in the small intestine via increased excretion of a mutagen-bacteria complex.