• Title/Summary/Keyword: beta-function

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Inactivation of the DevS Histidine Kinase of Mycobacterium smegmatis by the Formation of the Intersubunit Disulfide Bond (Subunit 간의 disulfide 결합 형성에 의한 Mycobacterium smegmatis DevS histidine kinase의 불활성화)

  • Lee, Jin-Mok;Park, Kwang-Jin;Kim, Min-Ju;Ko, In-Jeong;Oh, Jeong-Il
    • Journal of Life Science
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    • v.20 no.6
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    • pp.853-860
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    • 2010
  • The DevSR two-component system is a major regulatory system involved in redox sensing in Mycobacterium smegmatis. The DevSR system consists of the DevS histidine kinase and its cognate DevR response regulator. When exposed to hypoxic conditions, the DevS histidine kinase is activated to phosphorylate the DevR response regulator, leading to the transcriptional activation of the DevR regulation. The ligand-binding state of the heme embedded in the N-terminal GAF domain of DevS determines the kinase activity of DevS. In this study, we demonstrated that the redox-responsive cysteine (C547) in the C-terminal kinase domain is involved in the redox-dependent control of DevS kinase activity. The formation of an intersubunit disulfide bond between the C547 residues in the presence of $O_2$ led to inactivation of DevS kinase activity. The reduction of the oxidized DevS with reductants such as $\beta$-mercaptoethanol and dithiothreitol resulted in the restoration of DevS kinase activity. It was demonstrated in vivo by complementation test that the substitution of C547 to alanine partially impaired the sensory function of DevS in M. smegmatis.

Effect of Alpina Officinarum Ethanol Extract on Immunoregulatory Activities in the Mice (양강 에탄올 추출물이 마우스에서 면역조절작용에 미치는 영향)

  • Kim, Hyang Suk;Chung, Kyung Tae;Lee, In Hwan;Choi, Woo Bong;Lee, Jong Hwan;Hyun, Sook Kyung;Kim, Byung Woo;Hwang, Hye Jin
    • Journal of Life Science
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    • v.24 no.1
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    • pp.61-66
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    • 2014
  • The purpose of this study was to investigate the immunomodulatory effects of Alpina officinarum (AO) ethanol extract on immunocompromised mice. The mice were injected intraperitoneally with an immunosuppressive drug, cyclophosphamide, and then administrated orally with 30, 100, and 300 mg/kg of ethanol extract of AO (AO 30, AO 100, and AO 300, respectively). The concentrations of cytokines and immunoglobulins (IgM, IgA, IgG) in serum were measured. The body weight of the mice and spleen cell number of the AO-fed group showed no significant difference compared to a control group. The concentrations of several cytokines, including IL-2, IFN-${\gamma}$, and TGF-${\beta}$, in serum showed a significant increase in the AO 100 group compared to the control and other groups (p<0.05). The IL-4 level showed no significant difference in the experimental groups. The supplementation of AO (30, 100, 300 mg/kg) significantly increased the concentration of IgM (p<0.05). The concentration of IgA was significantly increased in the AO 100 group (p<0.05) compared to the control group. It can be concluded that AO ethanol extract enhances immune function by promoting the production of cytokines and immunoglobulins.

Circadian Clock Gene Per1 Mediates BMP2-induced Osteoblast Differentiation in MC3T3-E1 Cells (MC3T3-E1 세포에서 BMP2에 의한 조골세포의 분화에 일주기 유전자 Per1이 미치는 영향)

  • Min, Hyeon-Young;Jang, Won-Gu
    • Journal of Life Science
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    • v.27 no.5
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    • pp.501-508
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    • 2017
  • Bone morphogenetic proteins (BMPs) are multifunctional cytokines that play important roles in a variety of cellular functions. Among BMP family members, BMP2 efficiently promotes osteoblast differentiation through Smad-mediated runt-related transcription factor 2 (Runx2) expression. Several recent studies suggest that BMPs are associated with clock genes, in particular Bmal1. Bmal1 protein heterodimerizes with Clock protein and then induces period 1 (Per1) expression. However, the role of Per1 on osteoblast differentiation remains unclear. In this study, we investigated whether Per1 is involved in osteoblast differentiation. MC3T3-E1 cells were treated with BMP2 for induction of osteoblastic differentiation. Osteogenic maker gene and Per1 mRNA expression were measured using real-time PCR. Interestingly, BMP2 treatment induced Per1 mRNA expression in MC3T3-E1 cells. To further investigate the function of Per1 on osteoblast differentiation, MC3T3-E1 cells were transiently transfected with pCMV-Per1. Per1 overexpression increased Runx2 mRNA and protein levels. Also, mRNA expression and promoter activity of osteocalcin were upregulated by Per1 overexpression. To investigate the effect of interaction between Per1 and osteogenic condition, MC3T3-E1 cells were cultured in osteogenic medium containing ascorbic acid and ${\beta}$-glycerophosphate. Osteogenic medium-induced ALP staining level and mineralization were synergistically increased by overexpression of Per1. Taken together, these results demonstrate that Per1 is a positive regulator of osteoblast differentiation.

Anti-diabetic mechanism of melania snail (Semisulcospira libertina) protamex hydrolysates (다슬기 protamex 가수분해물(MPH)의 항당뇨 기작 연구)

  • Pyo, Sang-Eun;Choi, Jae-Suk;Kim, Mi-Ryung
    • Food Science and Preservation
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    • v.24 no.7
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    • pp.1007-1016
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    • 2017
  • Melania snail (Semisulcospira libertina) was traditionally used as the healthy food in Korea. It was generally known to improve liver function and heal a diabetes. The aim of this study was to elucidate the anti-diabetic mechanism of melanian snail hydrolysates treated with protamex (MPH) by investigating the inhibitory action on protein tyrosine phosphatase 1B (PTP1B), the improving effect on the insulin resistance in C2C12 myoblast and the protective effect for pancreatic beta-cell (INS-1) under the glucose toxicity. The melania snail hydrolysates treated with protamex (MPH), which showed the highest degree of hydrolysis (43%), and inhibited effectively PTP1B activity ($IC_{50}=15.42{\pm}1.1{\mu}g/mL$), of which inhibitory effect was higher than usolic acid, positive control ($IC_{50}=16.65{\mu}g/mL$). MPH increased the glucose uptake in C2C12 myoblast treated with palmitic acid. In addition, MPH increased insulin mRNA expression level by over 160% with enhanced cell viability in INS-1 cell under the high glucose concentration (30 mM). These results suggest that MHP may improve the diabetic symptom by the inhibiting the PTP1B activity, increasing the glucose uptake in muscle cell and protecting the pancreatic beta-cell from glucose toxicity.

Characteristics of White Soybean Chungkookjang Fermented by Bacillus subtilis D7 (Bacillus subtilis D7에 의하여 발효된 백태 청국장의 특성)

  • Lee, Na-Ri;Park, Sung-Bo;Lee, Sang-Mee;Go, Tae-Hun;Hwang, Dae-Youn;Kim, Dong-Seob;Jeong, Seong-Yun;Son, Hong-Joo
    • Journal of Life Science
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    • v.23 no.4
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    • pp.529-536
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    • 2013
  • This study was carried out to investigate the characteristics of white soybean Chungkookjang fermented by Bacillus subtilis D7. The highest germination rate was obtained under $25^{\circ}C$ when water was supplied for 4 days at intervals of 2 hr. The total isoflavone content was 971.3 ${\mu}g/g$ before germination and 1023.8 ${\mu}g/g$ after germination. The amino type- and ammonia type-nitrogen contents of Chungkookjang were proportional to the fermentation time. The pH values of all Chungkookjang soybeans increased up to pH 7.8-8.0 during fermentation. The number of viable cells in all Chungkookjang soybeans increased significantly up to 24 hr. The protease activity of all Chungkookjang soybeans increased up to 30-36 hr. The ${\alpha}$- and ${\beta}$-amylase activities of Chungkookjang fermented by B. subtilis D7 were enhanced with increasing fermentation time. Analysis of the effect of Chungkookjang intake on the liver function of mouse showed that the alkaline phosphatase (ALP) activity and the superoxide dismutase (SOD) activity in the Chungkookjang diet group were markedly higher than those in the control group. The asparatate aminotransferase (AST) activity in the germinated soybean Chungkookjang diet group was higher than that in the nongerminated soybean Chungkookjang diet group. Therefore, Chungkookjang soybeans fermented with B. subtilis D7 can be expected to have an increased content of functional components and improved quality characteristics.

Molecular Cloning, Sequence Analysis, and in Vitro Expression of Flavanone 3β-Hydroxylase from Gypsophila paniculata (안개초(Gyposphila paniculata)로부터 Flavanone 3β-Hydroxylase 유전자의 분리 및 분석)

  • Min, Byung-Whan
    • Journal of Plant Biotechnology
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    • v.33 no.2
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    • pp.85-91
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    • 2006
  • Flavanone 3$\beta$-hydroxylase (FHT) is an enzyme acting in the central part of the flavonoid biosynthesis pathway. FHT catalyses the hydroxylation of flavanone to dihydroflavonols in the anthocyanin pathway. In this paper we describe the cloning and expression of the genes encoding the flavonoid-biosynthetic enzyme FHT in Gypsophila paniculata L. A heterologous cDHA probe from Dianthus cavophyllus was used to isolate FHT-encoding cDHA clones from Gypsophila paniculata L.. Inspection of the 1471 bp long sequence revealed an open reading frame 1047 bp, including a 190 bp 5' leader region and 288 bp 3' untranslated region. Comparison of the coding region of this FHT cDHA sequence including the sequences of Arabidopsis thaliana, Citrus sinensis, Dianthus caryophyllus, Ipomoea batatas, Matthiola incana, Nierembergia sp, Petunia hybrida, Solanum tuberosum, Vitis vinifera reveals a identity higher than 69% at the nucleotide level. The function of this nucleotide sequences were verified by comparison with amino acid sequences of the amino-terminus and tryptic peptides from purified plant enzyme, by northern blotting with mRHA from wild type and mutant plants, by in vitro expression yielding and enzymatically active hydroxylase, as indicated by the small dihydrokaempferol peak. Genomic southern blot analysis showed the presence of only one gene for FHT in Gypsophila paniculata.

The Design of Feature Selection Classifier based on Physiological Signal for Emotion Detection (감성판별을 위한 생체신호기반 특징선택 분류기 설계)

  • Lee, JeeEun;Yoo, Sun K.
    • Journal of the Institute of Electronics and Information Engineers
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    • v.50 no.11
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    • pp.206-216
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    • 2013
  • The emotion plays a critical role in human's daily life including learning, action, decision and communication. In this paper, emotion discrimination classifier is designed to reduce system complexity through reduced selection of dominant features from biosignals. The photoplethysmography(PPG), skin temperature, skin conductance, fontal and parietal electroencephalography(EEG) signals were measured during 4 types of movie watching associated with the induction of neutral, sad, fear joy emotions. The genetic algorithm with support vector machine(SVM) based fitness function was designed to determine dominant features among 24 parameters extracted from measured biosignals. It shows maximum classification accuracy of 96.4%, which is 17% higher than that of SVM alone. The minimum error features selected are the mean and NN50 of heart rate variability from PPG signal, the mean of PPG induced pulse transit time, the mean of skin resistance, and ${\delta}$ and ${\beta}$ frequency band powers of parietal EEG. The combination of parietal EEG, PPG, and skin resistance is recommendable in high accuracy instrumentation, while the combinational use of PPG and skin conductance(79% accuracy) is affordable in simplified instrumentation.

Study on the Micellization of CPC/Brij 35 Mixed Surfactant Systems in Water (순수 물에서 CPC/Brij 35 혼합계면활성제의 미셀화에 대한 연구)

  • Gil, Han-Nae;Lee, Byung-Hwan
    • Journal of the Korean Chemical Society
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    • v.53 no.2
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    • pp.118-124
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    • 2009
  • The values of critical micelle concentration (CMC) and counter ion binding constants (B) in a micellar state of CPC (1-hexadecylpyridinium chloride) with Brij 35 (polyoxyethylene(23) lauryl ether) in water were determined as a function of ${\alpha}_1$ (the overall mole fraction of CPC) by the use of electric conductivity method. Various thermodynamic parameters ($X_i,\;{\gamma}_i,\;C_i,\;a_{i}^{M},\;\beta,\;and\;{\Delta}H_{mix}$) were calculated and analyzed by means of the equations derived from the non-ideal mixed micellar model. And thermodynamic parameters (${\Delta}{G^o}_m,\;{\Delta}{H^o}_m,\;and\;{\Delta}{S^o}_m$) for the micellization of CPC/Brij 35 mixtures were also calculated from the temperature dependence of the CMC values. The values of ${\Delta}{G^o}_m$ are all negative, but the values of ${\Delta}{S^o}_m$ and ${\Delta}{H^o}_m$ are positive or negative, depending on the measured temperature and ${\alpha}_1$.

The Effects of Chungpyesagan-tang on the Recovery of Liver Function in Rat Injured by $CCl_4$ (청폐사간탕(淸肺瀉肝湯)이 $CCl_4$로 유발(誘發)된 휜쥐의 간손상(肝損傷)에 마치는 영향(影響))

  • Shin, Mee-Ran;Heo, Woon-Yeong;Kim, DaI-Rae;Jeon, Jong-Weon;Kim, Jeong-Yeol
    • Journal of Sasang Constitutional Medicine
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    • v.17 no.1
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    • pp.130-141
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    • 2005
  • 1. Objectives The present study was carried out to investigate the effects of Chungpyesagan-tang on the $CCI_4-induced$ Liver Damage in Rats. 2. Methods Sprague-Dawley rats were devided into 5 experimental groups : Normal, $NS+CCI_4(Solid$ extract of $CCI_4$ injection group after Normal Saline feed), $CP+CCI_4(Solid$ extract of $CCI_4$ injection group after Chungpyesagantang feed), $CCI_4+NS(Nomal$ Saline feed group after $CCI_4$ injection), $CCI_4+CP(Solid$ extract of Chungpyesagantang feed group after $CCI_4$ injection). Biochemical assays for serum enzyme activities such as AST, ALT, ALP, BUN, Creatinine, Uric Acid, Total Protein, Albumin, Total Cholesterol, Triglyceride, Glucose, and mRNA Revelation of Cytochrome p450 and activities such as LPO(Lipid Peroxidation), GSH(Glutathione), GST(Glutathione-S- Transferase), Glutathione Reductase, Glutathione Peroxidase, SOD(Superoxide Dismutase), Catalase, Hydroxyproline, and ${\beta}-Glucuronidase$ were performed. 3. Results 1) $CP+CCI_4$ showed significantly lower relation of Cytochrome p450 than $NS+CCI_4$. 2) As to LPO Hydroxyproline, $CCI_4+CP$ showed significantly lower activity than $CCI_4+NS$. 3) As to GSH GST Glutathione Peroxidase Catalase, $CP+CCI_4$ showed higher activity than $NS+CCI_4$, $CCI_4+CP$ showed significantly higher activity than $CCI_4+NS$. 4) As to Glutathione Reductase SOD, $CCI_4+CP$ showed significantly higher activity than $CCI_4+NS$. 5) As to ${\beta}-Glucuronidase$, $CP+CCI_4$ showed signficantly lower activity than $NS+CCI_4$. 4. Conclusions Chungpyesagantang has the recovering effects on the $CCI_4-induced$ Liver Damage.

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Panax ginseng total protein promotes proliferation and secretion of collagen in NIH/3T3 cells by activating extracellular signal-related kinase pathway

  • Chen, Xuenan;Wang, Manying;Xu, Xiaohao;Liu, Jianzeng;Mei, Bing;Fu, Pingping;Zhao, Daqing;Sun, Liwei
    • Journal of Ginseng Research
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    • v.41 no.3
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    • pp.411-418
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    • 2017
  • Background: Recently, protein from ginseng was studied and used for the treatment of several kinds of diseases. However, the effect of ginseng total protein (GTP) on proliferation and wound healing in fibroblast cells remains unclear. Methods: In this study, cell viability was analyzed using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. Cell cycle distribution was analyzed by flow cytometer. The levels of transforming growth factor ${\beta}1$, vascular endothelial growth factor, and collagens were analyzed by enzyme-linked immunosorbent assay and immunofluorescence staining. The expressions of cyclin A, phosphorylation of extracellular signal-related kinase (p-ERK1/2), and ERK1/2 were analyzed by Western blotting. Results: Our results showed that GTP promoted cell proliferation and increased the percentage of cells in S phase through the upregulation of cyclin A in NIH/3T3 cells. We also found that GTP induced the secretion of type I collagen, and promoted the expression of other factors that regulate the synthesis of collagen such as transforming growth factor ${\beta}1$ and vascular endothelial growth factor. In addition, the phosphorylation of ERK1/2 at Thr202/Tyr204 was also increased by GTP. Conclusion: Our studies suggest that GTP promoted proliferation and secretion of collagen in NIH/3T3 cells by activating the ERK signal pathway, which shed light on a potential function of GTP in promoting wound healing.