• Title/Summary/Keyword: metB

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Biochemical Analysis on the Parallel Pathways of Methionine Biosynthesis in Corynebacterium glutamicum

  • Hwang, Byung-Joon;Park, Soo-Dong;Kim, Youn-Hee;Kim, Pil;Lee, Heung-Shick
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.1010-1017
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    • 2007
  • Two alternative pathways for methionine biosynthesis are known in Corynebacterium glutamicum: one involving transsulfuration (mediated by metB and metC) and the other involving direct sulfhydrylation (mediated by metY). In this study, MetB (cystathionine ${\gamma}-synthase$) and MetY (O-acetylhomoserine sulfhydrylase) from C. glutamicum were purified to homogeneity and the biochemical parameters were compared to assess the functional and evolutionary importance of each pathway. The molecular masses of the native MetB and MetY proteins were measured to be approximately 170 and 280 kDa, respectively, showing that MetB was a homotetramer of 40-kDa subunits and MetY was a homohexamer of 45-kDa subunits. The $K_m$ values for the O-acetylhomoserine catalysis effected by MetB and MetY were 3.9 and 6.4 mM, and the maximum catalysis rates were $7.4\;(k_{cat}=21\;S^{-1})\;and\;6.0\;(k_{cat}=28\;S^{-1})\;{\mu}mol\;mg^{-1}\;min^{-1}$, respectively. This suggests that both MetB and MetY can be comparably active in vivo. Nevertheless, the $K_m$ value for sulfide ions by MetY was 8.6mM, which was too high, considering the physiological condition. Moreover, MetB was active at a broad range of temperatures $(30\;and\;65^{\circ}C)$ and pH (6.5 and 10.0), as compared with MetY, which was active in a range from 30 to $45^{\circ}C$ and at pH values from 7.0 to 8.5. In addition, MetY was inhibited by methionine, but MetB was not. These biochemical data may provide insight on the role of the parallel pathways of methionine biosynthesis in C. glutamicum with regard to cell physiology and evolution.

The Clinical Research of the Effectiveness of Muscle Energy Technique (MET) on Peripheral Facial Paralysis

  • Jo, Na Young;Roh, Jeong Du
    • The Journal of Korean Medicine
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    • v.37 no.2
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    • pp.45-52
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    • 2016
  • Objectives: The purpose of this study is to evaluate the clinical effects of Muscle Energy Technique(MET) for peripheral facial paralysis. Methods: 60 Patients were divided into two groups. Group A(n=30) received the treatment with existing Korean medicine. Group B(n=30) received the MET with existing Korean medicine. It was performed once a day, five time per a week for three weeks. we analyzed Yanagihara's score and House-Brackmann scale Results: A week after MET treatment, Yanagihara's score average of Group A is $7.17{\pm}6.34$. Yanagihara's score average of Group B is $8.84{\pm}5.22$. (p=0.72). Two weeks after MET, Yanagihara's score average of Group A is $12.39{\pm}4.94$. Yanagihara's score average of Group B is $15.12{\pm}3.20$. (p=0.04). Three weeks after MET, Yanagihara's score average of Group A is $17.11{\pm}5.31$. Yanagihara's score average of Group B is $22.78{\pm}3.67$. (p=0.01). A is $3.87{\pm}1.36$. House-Brackmann Scale average of Group B is $3.64{\pm}1.76$. (p=0.63). Two weeks after MET treatment, House-Brackmann Scale average of Group A is $3.20{\pm}0.97$. House-Brackmann Scale average of Group B is $3.02{\pm}1.03$. (p=0.05). Three weeks after MET, House-Brackmann Scale average of Group A is $2.84{\pm}1.12$. House-Brackmann Scale average of Group B is $2.23{\pm}0.78$. (p=0.04). Conclusion: MET treatment is effective for improve the symptoms of peripheral facial paralysis. Therefore, it will be used to peripheral facial paralysis.

Validation of Satellite Scatterometer Sea-Surface Wind Vectors (MetOp-A/B ASCAT) in the Korean Coastal Region (한반도 연안해역에서 인공위성 산란계(MetOp-A/B ASCAT) 해상풍 검증)

  • Kwak, Byeong-Dae;Park, Kyung-Ae;Woo, Hye-Jin;Kim, Hee-Young;Hong, Sung-Eun;Sohn, Eun-Ha
    • Journal of the Korean earth science society
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    • v.42 no.5
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    • pp.536-555
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    • 2021
  • Sea-surface wind is an important variable in ocean-atmosphere interactions, leading to the changes in ocean surface currents and circulation, mixed layers, and heat flux. With the development of satellite technology, sea-surface winds data retrieved from scatterometer observation data have been used for various purposes. In a complex marine environment such as the Korean Peninsula coast, scatterometer-observed sea-surface wind is an important factor for analyzing ocean and atmospheric phenomena. Therefore, the validation results of wind accuracy can be used for diverse applications. In this study, the sea-surface winds derived from ASCAT (Advanced SCATterometer) mounted on MetOp-A/B (METeorological Operational Satellite-A/B) were validated compared to in-situ wind measurements at 16 marine buoy stations around the Korean Peninsula from January to December 2020. The buoy winds measured at a height of 4-5 m from the sea surface were converted to 10-m neutral winds using the LKB (Liu-Katsaros-Businger) model. The matchup procedure produced 5,544 and 10,051 collocation points for MetOp-A and MetOp-B, respectively. The root mean square errors (RMSE) were 1.36 and 1.28 m s-1, and bias errors amounted to 0.44 and 0.65 m s-1 for MetOp-A and MetOp-B, respectively. The wind directions of both scatterometers exhibited negative biases of -8.03° and -6.97° and RMSE values of 32.46° and 36.06° for MetOp-A and MetOp-B, respectively. These errors were likely associated with the stratification and dynamics of the marine-atmospheric boundary layer. In the seas around the Korean Peninsula, the sea-surface winds of the ASCAT tended to be more overestimated than the in-situ wind speeds, particularly at weak wind speeds. In addition, the closer the distance from the coast, the more the amplification of error. The present results could contribute to the development of a prediction model as improved input data and the understanding of air-sea interaction and impact of typhoons in the coastal regions around the Korean Peninsula.

Effects of Epothilone A in Combination with the Antidiabetic Drugs Metformin and Sitagliptin in HepG2 Human Hepatocellular Cancer Cells: Role of Transcriptional Factors NF-κB and p53

  • Rogalska, Aneta;Sliwinska, Agnieszka;Kasznicki, Jacek;Drzewoski, Jozef;Marczak, Agnieszka
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.3
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    • pp.993-1001
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    • 2016
  • Type 2 diabetes mellitus patients are at increased risk of many forms of malignancies, especially of the pancreas, colon and hepatocellular cancer. Unfortunately, little is known of the possible interaction between antidiabetic drugs and anticancer agents. The present study investigates the influence of metformin (MET) and sitagliptin (SITA) on the in vitro anticancer activity of the microtubule depolymerization inhibitor agent epothilone A (EpoA). Hepatocellular liver carcinoma cell line (HepG2) viability and apoptosis were determined by the MTT test and by double staining with PO-PRO-1 and 7-aminoactinomycin D, respectively, after treatment with EpoA, metformin or sitagliptin. The levels of nuclear factor NF-${\kappa}B$ and p53 were evaluated in the presence and absence of inhibitors. While EpoA and MET inhibited HepG2 cell proliferation, SITA did not. EpoA and SITA induced higher p53 levels than MET. All tested drugs increased the level of NF-${\kappa}B$. Only MET enhanced the proapoptotic effect of EpoA. The EpoA+MET combination evoked the highest cytotoxic effect on HepG2 cells and led to apoptosis independent of p53, decreasing the level of NF-${\kappa}B$. These findings support the link between NF-${\kappa}B$ and p53 in the modulation of apoptotic effects in HepG2 cells treated by EpoA. Our studies indicate that the combination of EpoA and MET applied in subtoxic doses has a stronger cytotoxic effect on liver cancer cells than each of the compounds alone. The therapeutic advantages of the combination of EpoA with MET may be valuable in the treatment of patients with diabetes mellitus type 2 (T2DM) and liver cancer.

Cytoplasmic Inheritance of High Tillering and Earing Characters of a Korean Local Maize Line(MET) (다수.다얼성 옥수수(NET)의 세포질적 유전)

  • Han, C.H.;Lee, I.S.;Choe, B.H.;Park, K.Y.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.29 no.1
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    • pp.55-61
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    • 1984
  • A Korean local maize line, MET, which has multi-ears and tillers has been proved as a potential source for silage production. However, no fundamental genetic nature for the line has been investigated. Therefore, this study was done to find genetic information on the multi-earing and -tillering habits of MET line. MET line and a hybrid. (Mo 17 ${\times}$ B68), with monoculm and single ear per plant were used for production of F$_1$(F$\_$1-12/ and F$\_$1-21/), F$\_$2-12/, F$\_$2-21/, BC$\_$1-12/ and BC$\_$1-21/ generations. From the comparison of reciprocal crosses, it was found that the tillering and earing habits of the MET line are controlled by cytoplasmic factors. The tiller and ear numbers, and barren ears were all characters associated with the MET cytoplasm. The cytoplasmic effect of MET on tiller and ear numb en was not evident in F$_1$ generation, probably because of suppressing effect of heterosis on appearance of tillers or ears. Genetic parameters for the gene action for both tiller and ear number also indicated a lack of mono- or digenic-chromosomal gene effects. The heritability (broad) was very low for both characters. Therefore, it is strongly concluded that the tillering and earing characters of MET line are due to cytoplasmic reasons.

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Relative Performance and Immune Response in White Leghorn Layers Fed Liquid DL-methionine Hydroxy Analogue and DL-methionine

  • Panda, A.K.;Rama Rao, S.V.;Raju, M.V.L.N.;Bhanja, S.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.6
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    • pp.948-953
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    • 2007
  • The relative performance and immune response was evaluated in White Leghorn layers fed liquid DL-methionine hydroxyl analogue-free acid (MHA-FA) relative to dry DL-methionine (DLM) in maize-soybean-sunflower based diets. Three graded levels of methionine (Met) from DLM or MHA-FA were added to the basal diet containing 0.27% Met on an equimolar basis to achieve 0.30, 0.36 and 0.42% Met in the diet. Each diet was fed ad libitum to 25 replicates of one bird (individual feeding) each, from 24 to 40 weeks of age. A regime of 16 h light was provided and all the layers were kept under uniform management throughout the experimental period. None of the parameters studied were influenced by the interaction between source and level of Met in diets. Similarly, the majority of parameters, except for daily feed consumption and immune response (influenced by level) and egg specific gravity and shell thickness (influenced by source), were not affected by either source or level of Met in the diets. Feed consumption was significantly lower in the birds fed a diet containing 0.42% Met compared to those fed lower levels of Met. The cutaneous basophilic hypersensitivity response to PHA-P and antibody titre (32 and 40 wk) to inoculation of sheep red blood cells increased significantly by increasing the concentration of Met in the diet from 0.30 to 0.36%. Thus, the Met requirement for immune competence was higher than for optimum production. The source of Met significantly influenced the egg specific gravity and shell thickness. The specific gravity and shell thickness of eggs increased significantly when MHA-FA was used as the source of Met in the diet compared to DLM. From the study it is concluded that Met requirement for immune competence (360 mg/b/d) is higher than for optimum production (300 mg/b/d). MHA-FA was comparable with DLM as a source of Met for production performance and immunity, when the bioavailability of MHA-FA was considered as 88% of DLM. Further, MHA-FA improved egg shell quality compared to DLM.

Association between dietary flavanones intake and lipid profiles according to the presence of metabolic syndrome in Korean women with type 2 diabetes mellitus

  • Oh, Ji Soo;Kim, Hyesook;Vijayakumar, Aswathy;Kwon, Oran;Choi, Young Ju;Huh, Kap Bum;Chang, Namsoo
    • Nutrition Research and Practice
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    • v.10 no.1
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    • pp.67-73
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    • 2016
  • BACKGROUND/OBJECTIVES: This study was aimed at examining the association between dietary flavanones intake and lipid profiles according to the presence of metabolic syndrome (MetS) in Korean women with type 2 diabetes mellitus (T2DM). SUBJECTS/METHODS: A cross-sectional analysis was performed among 502 female T2DM patients (non-MetS group; n = 129, MetS group; n = 373) who were recruited from the Huh's Diabetes Clinic in Seoul, Korea between 2005 and 2011. The dietary intake was assessed by a validated semi-quantitative food frequency questionnaire (FFQ) and the data was analyzed using the Computer Aided Nutritional Analysis program (CAN-Pro) version 4.0 software. The intake of flavanones was estimated on the basis of the flavonoid database. RESULTS: In the multiple linear regression analysis after adjustment for confounding factors, daily flavanones intake was negatively associated with CVD risk factors such as total cholesterol, LDL-cholesterol, and apoB and apoB/apoA1 ratio only in the MetS group but not in the non-MetS group. Multiple logistic regression analysis revealed that the odds ratio for a higher apoB/apoA1 ratio above the median (${\geq}0.74$) was significantly low in the $4^{th}$ quartile compared to that in the $1^{st}$ quartile of dietary flavanones intake [OR: 0.477, 95% CI: 0.255-0.894, P for trend = 0.0377] in the MetS group. CONCLUSIONS: Dietary flavanones intake was inversely associated with the apoB/apoA1 ratio, suggesting a potential protective effect of flavanones against CVD in T2DM women with MetS.

Construction and Characterization of Vector Expressing Low Level of Translation Factor eIF5B (단백질합성인자 eIF5B의 저 발현 효모벡터의 제조 및 특성)

  • 최상기;송진희;이준행;이병욱;성치남
    • Korean Journal of Microbiology
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    • v.40 no.1
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    • pp.7-11
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    • 2004
  • eIF5B is a translation initiation factor that delivers Met-$tRNA^{Met}$ to AUG start codon and subsequently joins the small and large ribosomes. In order to study the function of eIF5B encoded by FUN12, we constructed FUN12 which lacked 5' end of its sequence. We found that this construct lacking almost all of its promoter in pRS plasmid partially complemented slow growth phenotype of fun12 deletion strain. Interestingly, this construct expressed N-terminally truncated eIF5B and its expression level was about 5% of that of wild type eIF5B. Low amount of the eIF5B expressed additionally in fun12 deletion strain played a direct role as a limiting factor for its growth. This limiting factor eIF5B in those strains also modulates activities of overall translation in vitro.

Regulation of Enzymes Involved in Methionine Biosynthesis in Corynebacterium glutamicum

  • Yeom, Hye-Jin;Hwang, Byung-Joon;Lee, Myong-Sok;Kim, Youn-Hee;Lee, Heung-Shick
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.373-378
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    • 2004
  • The regulatory mechanism of methionine biosynthesis in Corynebacterium glutamicum was analyzed at the protein arid gene expression level. O-Acetylhomoserine sulfhydraylase (encoded by metY) was inhibited by 10 mM methionine to a residual activity of 10% level, whereas no such inhibition was found with cystathionine $\gamma$-synthase (encoded by metB) and cystathionine $\beta$-lyase (encoded by metC). The enzymatic activity of homoserine acetyltransferase (encoded by metX) was repressed to a residual activity of 25% level by 10 mM methionine which was added to the growth medium. Cystathionine $\gamma$-synthase and cystathionine $\beta$-lyase were also repressed by 10 mM methionine, but only to a residual activity of 50-70% level. O-Acetylhomoserine sulfhydrylase was very sensitive to repression by 10 mM methionine, showing residual activity of 13%. In addition, homoserine acetyltransferase was also repressed by 10 mM cysteine to 50% of its original activity. No repression of the enzymes by S-adenosyl methionine was observed. The pattern of repression by methionine indicated that the metB and aecD genes might be regulated by a common mechanism, while the metA and metY genes are differently regulated.

Enhanced supply of methionine regulates protein synthesis in bovine mammary epithelial cells under hyperthermia condition

  • Zhou, Jia;Yue, Shuangming;Xue, Benchu;Wang, Zhisheng;Wang, Lizhi;Peng, Quanhui;Xue, Bai
    • Journal of Animal Science and Technology
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    • v.63 no.5
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    • pp.1126-1141
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    • 2021
  • Recent evidence has shown that methionine (Met) supplementation can improve milk protein synthesis under hyperthermia (which reduces milk production). To explore the mechanism by which milk protein synthesis is affected by Met supplementation under hyperthermia, mammary alveolar (MAC-T) cells were incubated at a hyperthermic temperature of 42℃ for 6 h in media with different concentrations of Met. While the control group (CON) contained a normal amino acid concentration profile (60 ㎍/mL of Met), the three treatment groups were supplemented with Met at concentrations of 10 ㎍/mL (MET70, 70 ㎍/mL of Met), 20 ㎍/mL (MET80, 80 ㎍/mL of Met), and 30 ㎍/mL (MET90,90 ㎍/mL of Met). Our results show that additional Met supplementation increases the mRNA and protein levels of BCL2 (B-cell lymphoma-2, an anti-apoptosis agent), and decreases the mRNA and protein levels of BAX (Bcl-2-associated X protein, a pro-apoptosis agent), especially at an additional supplementary concentration of 20 ㎍/mL (group Met80). Supplementation with higher concentrations of Met decreased the mRNA levels of Caspase-3 and Caspase-9, and increased protein levels of heat shock protein (HSP70). The total protein levels of the mechanistic target of rapamycin (mTOR) and the mTOR signalling pathway-related proteins, AKT, ribosomal protein S6 kinase B1 (RPS6KB1), and ribosomal protein S6 (RPS6), increased with increasing Met supplementation, and peaked at 80 ㎍/mL Met (group Met80). In addition, we also found that additional Met supplementation upregulated the gene expression of αS1-casein (CSN1S1), β-casein (CSN2), and the amino acid transporter genes SLC38A2, SLC38A3 which are known to be mTOR targets. Additional Met supplementation, however, had no effect on the gene expression of κ-casein (CSN3) and solute carrier family 34 member 2 (SLC34A2). Our results suggest that additional Met supplementation with 20 ㎍/mL may promote the synthesis of milk proteins in bovine mammary epithelial cells under hyperthermia by inhibiting apoptosis, activating the AKT-mTOR-RPS6KB1 signalling pathway, and regulating the entry of amino acids into these cells.