• Title/Summary/Keyword: $G{\beta}{\gamma}$

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Purification and Characterization of Cyclodextrinase from Bacillus stearothermophilus KJ 16 (Bacillus stearothermophilus KJ16이 생산하는 Cyclodextrinase의 정제와 효소특성)

  • 권현주;유동주;김병우
    • Journal of Life Science
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    • v.8 no.5
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    • pp.497-503
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    • 1998
  • Cyclodextrinase from B. stearothemophilus KJ16 that can produce both cyclodextrin(CD) glucanotransferase and cyclodextrinase was purified 87.6-fold with 7% yield by ammonium sulfate precipitation, DEAE-cellulose chromatog-raphy, Sephadex G-100 chromatography, and FPLC. The molecular weight of the purified enzyme was about 68,000 dalton by SDS-PAGE. The optimal pH and temperature were 6.0 and 55$^{\circ}C$, respectively. The enzyme was stable at 5$0^{\circ}C$ for 2 hr in the pH range of 5.5 and 8.5. The enzyme activity was inhibited strongly by mercaptoethanol, di-thiothreitol, p-chloromercuribenzoate, N-bromosuccinimide, $Cu^{+2}$and $Hg^{+2}$. The purified enzyme hydrolyzed CDs with$\gamma$-CD>$\beta$-CD>$\alpha$-CD. The enzyme also hydrolyzed linear maltodextrins and polysaccharides, but the rates of hyd-rolysis for such substrates were slow as compared to that for $\gamma$-CD. The final degradation products with all substrates were maltose and glucose. Maltose was not further hydrolyzed.

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Tiul1 and TGIF are Involved in Downregulation of $TGF{\beta}1$-induced IgA Isotype Expression

  • Park, Kyoung-Hoon;Nam, Eun-Hee;Seo, Goo-Young;Seo, Su-Ryeon;Kim, Pyeung-Hyeun
    • IMMUNE NETWORK
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    • v.9 no.6
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    • pp.248-254
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    • 2009
  • [ $TGF-{\beta}1$ ]is well known to induce Ig germ-line ${\alpha}$ ($GL{\alpha}$) transcription and subsequent IgA isotype class switching recombination (CSR). Homeodomain protein TG-interacting factor (TGIF) and E3-ubiquitin ligases TGIF interacting ubiquitin ligase 1 (Tiul1) are implicated in the negative regulation of $TGF-{\beta}$ signaling. In the present study, we investigated the roles of Tiul1 and TGIF in $TGF{\beta}1$-induced IgA CSR. We found that over-expression of Tiul1 decreased $TGF{\beta}1$-induced $GL{\alpha}$ promoter activity and strengthened the inhibitory effect of Smad7 on the promoter activity. Likewise, overexpression of TGIF also diminished $GL{\alpha}$ promoter activity and further strengthened the inhibitory effect of Tiul1, suggesting that Tiul1 and TGIF can down-regulate $TGF{\beta}1$-induced $GL{\alpha}$ expression. In parallel, overexpression of Tiul1 decreased the expression of endogenous IgA CSR-predicitive transcripts ($GLT_{\alpha},\;PST_{\alpha},\;and\;CT_{\alpha}$) and $TGF{\beta}1$-induced IgA secretion, but not $GLT_{\gamma3}$ and IgG3 secretion. Here, over-expressed TGIF further strengthened the inhibitory effect of Tiul1. These results suggest that Tiul1 and TGIF act as negatively regulators in $TGF{\beta}1$-induced IgA isotype expression.

Extractive Nitrogenous Constituents and Their Monthly Variation of Fresh Laver Porphyra dentata (잇바디돌김의 함질소 엑스성분조성 및 월별변동)

  • 박춘규;박철훈;박정임
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.4
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    • pp.579-588
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    • 2001
  • In order to investigate the composition and monthly variation of extractive nitrogenous components in the laver Porphyra dentata cultured at the south coast of Korea, the fresh laver was analyzed separately for the amounts of free amino acids, combined amino acids, ATP and its related compounds, and quaternary ammonium basis in fresh laver were measured. The extractive nitrogen contents of fresh laver extracts were 760~870 mg/100g (dry basis). Twenty-seven to thirty-one kinds of free amino acids were detected in the laver extracts and their total amounts were 2,404~3,966 mg/100g (on dry basis). The laver extracts showed rich in free amino acids such as alanine, taurine, glutamic acid, glutamine and aspartic acid. Sixteen to twenty-three kinds of combined amino acids were detected in the extracts and their total amounts were 1,429~2,692 mg/100g (on dry basis). Proline, glutamic acid, glycine, phosphoserine, serine were the amin combined amino acids in the extracts. The amounts of ATP and related compounds were 73.3~94.4 mg/100 g (2.04~4.43 $\mu$mol/g, on dry basis). Homarine and trigonelline were detected in all specimens but $\beta$-alaninebetaine, ${\gamma}$-butyrobetaine were found in some. Small amounts of trimethylamine were detected in all samples. Free and combined amino acids were occupying almost 90% of extractive nitrogen. Most of free and combined amino acids showed a marked monthly variation with a maximum in January and March, and a minimum in February and April. The fresh laver P. dentata did not differ much from the fresh laver P. yezoensis in qualitative com-position of extractive components, but their contents were generally low level.

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Heterotrimeric G protein signaling and RGSs in Aspergillus nidulans

  • Yu Jae-Hyuk
    • Journal of Microbiology
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    • v.44 no.2
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    • pp.145-154
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    • 2006
  • Heterotrimeric G proteins (G proteins) are conserved in all eukaryotes and are crucial components sensing and relaying external cues into the cells to elicit appropriate physiological and biochemical responses. Basic units of the heterotrimeric G protein signaling system include a G protein-coupled receptor (GPCR), a G protein composed of ${\alpha},\;{\beta},\;and\;{\gamma}$ subunits, and variety of effectors. Sequential sensitization and activation of these G protein elements translates external signals into gene expression changes, resulting in appropriate cellular behaviors. Regulators of G protein signaling (RGSs) constitute a crucial element of appropriate control of the intensity and duration of G protein signaling. For the past decade, G protein signaling and its regulation have been intensively studied in a number of model and/or pathogenic fungi and outcomes of the studies provided better understanding on the upstream regulation of vegetative growth, mating, development, virulence/pathogenicity establishment, and biosynthesis of secondary metabolites in fungi. This review focuses on the characteristics of the basic upstream G protein components and RGS proteins, and their roles controlling various aspects of biological processes in the model filamentous ascomycete fungus Aspergillus nidulans. In particular, their functions in controlling hyphal proliferation, asexual spore formation, sexual fruiting, and the mycotoxin sterigmatocystin production are discussed.

Serum Protein Values of Racehorse in Korea (한국경주마의 혈청단백질치)

  • Lee Chang-Woo;Lee Sung-Yub;Lee Sung-Mo;Eom Yeong-Ho
    • Journal of Veterinary Clinics
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    • v.4 no.2
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    • pp.445-448
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    • 1987
  • 임상적으로 건강하고 휴식상태에 있는 한국 경주마의 혈청단백질치를 조사하였다. 총단백질농도는 6.5$\pm$0.3g/100ml(평균$\pm$표준편차)이였다. 알부민 농도는 3.4$\pm$0.2, $\alpha$-글로부린 농도는 0.8$\pm$0.1, $\beta$-글로부린 농도는 1.2$\pm$0.2, ${\gamma}$-글로부린 농도는 1.1$\pm$0.3g/100ml이었으며, A/G 비는 1.15$\pm$0.21이었다. 성, 연령, 계절에 따르는 혈청단백질치의 차이를 통계적으로 분석하여 보고하였다.

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Physicochemical Properties of Domestic Cherry Tomato Varieties (국내산 방울토마토의 이화학적 특성)

  • Ahn, Jun-Bae
    • Culinary science and hospitality research
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    • v.23 no.7
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    • pp.42-49
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    • 2017
  • This study was conducted to determine the nutritional value of domestic cherry tomato varieties (Summerking, Qutiquti, and Minichal). The levels of amino acids, amino acid derivatives, and ${\gamma}-aminobutyric-acid$ (GABA) were analyzed using ion chromatography. In domestic cherry tomatoes, eighteen free amino acids were found including L-glutamic acid (L-Glu), L-glutamine (L-Gln), and L-aspartic acid (L-Asp). L-Glu was the most abundant amino acid, ranging from 1,533.17 mg/100 g to 1,920.65 mg/100 g (dry weight). The next abundant amino acids were L-Gln, ranging from 784.68 mg/100 g to 1,164.36 mg/100 g and L-Asp, ranging from 320.73 mg/100 g to 387.22 mg/100 g. Domestic cherry tomatoes contained eight essential amino acids except tryptophan and the total essential amino acid content was 297.30~432.43 mg/100 g (dry weight), which was 8.92~10.61% of total free amino acid. Several amino acid derivatives were found: L-carnitine (L-Car), hydroxylysine (Hyl), o-phosphoethanolamine (o-Pea), phosphoserine (p-Ser), ${\beta}-alanine$ (${\beta}-Ala$), N-methyl-histidine (Me-His), ethanolamine ($EtNH_2$), and L-citrulline (L-Cit). L-Car, transporting long-chain fatty acid into mitocondrial matrix, was the most abundant amino acid derivative in all domestic cherry tomatoes. A high level of GABA (313.18~638.57 mg/100 g), known as a neurotransmitter, was also found in all three domestic cherry tomatoes. These results revealed that domestic cherry tomatoes have a good balance of nutrient and bioactive compounds. Therefore, cherry tomatoes can be used as a functional food material.

Mixed Micellar Properties of DPC with Other Cationic Surfactants (DTAB, TTAB, and CDEAB) (DPC와 다른 양이온성 계면활성제(DTAB, TTAB 및 CDEAB)와의 혼합마이셀화에 관한 연구)

  • Lee, Byung Hwan
    • Journal of the Korean Chemical Society
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    • v.42 no.5
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    • pp.519-525
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    • 1998
  • The critical micelle concentration $(CMC^{\ast})$ and the counterion binding constant (B) in a micellar state of the mixed surfactant systems of dodecylpyridinium chloride (DPC) with dodecyltrimethylammonium bromide (DTAB), tetradecyltrimethylammonium bromide (TTAB), and cetyldimethylethylammonium bromide (CDEAB) were determined at $25^{\circ}C$ as a function of ${\alpha}_1$ (the overall molc fraction of DPC) by the use of electric conductivity method. Various thermodynamic parameters $(X_{i},\;{\gamma}_{I},\;C_{i},\;a^{M}_{i}, \beta,\;{\Delta}H_{mix}, \;and\; {\Delta}G^{o}_{m})$ for the micellization of DPC/DTAB, DPC/TTAB, and DPC/CDEAB mixtures were calculated and analyzed for each system by means of the equations derived from the pseudophase separation model. The results show that the DPC/DTAB mixed system has the greatest deviation and the DPC/CDEAB mixed system has the smallest deviation from the ideal mixed micellar model.

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Purification and Characterization of Fibrinolytic Enzyme from Tricholoma saponaceum (II) (할미송이버섯으로부터 혈전용해효소의 정제 및 특성 연구 (II))

  • 김준호
    • Biomedical Science Letters
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    • v.6 no.4
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    • pp.261-268
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    • 2000
  • Fibrinolytic enzyme (FE-2) was purified from the fruiting bodies of Tricholoma saponaceum using DEAE-Cellulose chromatography and Mono-S column chromatography, The enzyme has a molecular weight of 18.23 kDa and include Zn$^{2+}$ ion as found by ICP/MS. The N-terminal amino acid sequence of the enzyme was A-L-Y-V-G-X-S-P-X-Q-Q-S-L-L-V It has a pH optimum at pH 7.5, suggested that FE-2 was a neutral pretense. The activity of FE-2 was highly inhibited by EDTA and 1,10-phenanthroline, indicating that the enzyme is a metalloprotease. The activity of FE-2 was increased by $Mg^{2+}$, Zn$^{2+}$, Fe$^{2+}$, and Co$^{2+}$, but the enzyme activity was totally inhibited by Hg$^{2+}$. No inhibition was found with PMSF, E-64, pepstatin and 2-mercaptoethanol. The enzyme hydrolyzed both $A\alpha$ and B$\beta$ chains of human fibrinogen. The $\gamma$ chain was resistant to hydrolysis by FE-2.

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Effects of the Houttuyniae Herba Extract on the Membranous Nephropathy induced by Cationic Bovine Serum Albumin in Mice (어성초(魚腥草)가 Cationic Bovine Serum Albumin 투여로 유발된 Membranous Nephropathy Mouse Model에 미치는 영향)

  • Jung, Dae-Ho;Cho, Chung-Sik;Kim, Cheol-Jung
    • The Journal of Korean Medicine
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    • v.30 no.4
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    • pp.93-107
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    • 2009
  • Objective: Membranous nephropathy (MN) is one of the most common causes of nephrotic syndrome in adults. However, there is not a satisfactory treatment for MN. This study aimed to evaluate the effect of Houttuyniae Herba Extract (HHE) on MN induced by cationic bovine serum albumin (cBSA). Methods: Mice were divided into 4 groups. The first group, Normal, was injected with saline. The second group, Control, was treated with cBSA (10mg/kg i.p) only. The third group, HHE-250, was treated with cBSA (10mg/kg i.p) and HHE (250mg/kg, p.o). The fourth group, HHE-500, was treated with cBSA (10mg/kg i.p) and HHE (500mg/kg, p.o). After treatment for 4 weeks, we measured change of body weight, 24 hrs proteinuria, serum albumin, total cholesterol, triglyceride, BUN, creatinine, IgA, IgM, IgG, TNF-${\alpha}$, IL-1${\beta}$ levels and the mRNA expression of IFN-${\gamma}$, IL-6, and IL-10. The morphologic changes of renal glomeruli were also observed with a light microscope and an electron microscope. Results: The levels of 24 hrs proteinuria and serum triglyceride, BUN, IgG, TNF-${\alpha}$, IL-1${\beta}$ significantly decreased in both HHE groups, while the level of serum albumin significantly increased in both HHE groups. The mRNA expression of IFN-${\gamma}$ and IL-6 in splenocytes considerably increased in both HHE groups. The mRNA expression of IL-10 in splenocytes considerably decreased in both HHE groups. In histological findings of kidney tissue, thickening of GBM decreased in both HHE groups. Conclusions: This study shows that HHE might be effective for treatment of acute stage MN. More clinical data and studies are to be done for efficient application.

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Purification and Characterization of Cyclodextrin Glycosyltransferase from Bacillus brevis CD162 (Bacillus brevis CD162 Cyclodextrin Glycosyltransferase의 정제 및 특성)

  • Kim, Myung-Hee;Lim, Young-Hee;Oh, Tae-Kwang;Sohn, Cheon-Bae
    • Applied Biological Chemistry
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    • v.40 no.6
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    • pp.465-471
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    • 1997
  • The cyclodextrin glycosyltransferase (CGTase, EC 3.2.1.19) from Bacillus brevis CD162 was purified by precipitating with ammonium sulfate, DEAE-Sepharose CL-6B column chromatography and Sephadex G-150 column chromatography. The molecular mass and pI of the purified enzyme were estimated to be 74,000 and 6.3 by SDS-PAGE and isoelectric focusing, respectively. The purified enzyme was clearly identified as the CGTase by zymogram after SDS-PAGE. The optimum pH and temperature for the enzyme activity were 8.0 and $55^{\circ}C$, respectively. The enzyme was stable at the range of pH $5.5{\sim}9.0$, and up to $50^{\circ}C$. The amino acid sequence from the $NH_2-terminal$ of the purified CGTase was Ser-Val-Thr-Asn-Lys-Val-Asn-Tyr-Ser-Lys-Asp-Val-Ile-Tyr-Gln. The yields of the products from starch as the substrate were 1.3% for ${\alpha}-$, 33.9% for ${\beta}-$, and 9.7% for ${\gamma}-cyclodextrin$.

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