• Title/Summary/Keyword: ${\beta}-2$ protein

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Effect of IPTG Induction on Production of ${\beta}$-Galactosidase-PreS2 Fusion Protein in Recombinant Escherichia coli

  • Nam, Soo-Man;Park, Young-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.1 no.4
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    • pp.274-280
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    • 1991
  • Effects of IPTG induction on cell growth and production of ${\beta}$-galactosidase-preS2 fusion protein (${\beta}$gal-preS2) were studied in a defined medium using a recombinant Escherichia coli JM109/pCMHB30. IPTG was added (0.2 mM) to induce the cloned-gene expression in the early-, mid-, and late-log growth phases. The most serious decreases in growth rate and plasmid stability were observed for the induction in the early-log growth phase. The expression level of ${\beta}$gal-preS2 attained by the induction in the mid-log phase was about 0.51 mg fusion protein/mg total cellular protein, which was 2- and 5-fold improvement over the levels obtained with the inductions in the early- and late-log phases. Formation of acidic byproducts including acetate and pyruvate showed different profiles during the fermentation period for each cases of induction; pyruvate was the major byproduct for the induction in the early-log phase while acetate production became more significant for the cases of inductions in the mid- and late-log phases.

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Moutan Cortex Extract Inhibits Amyloid ${\beta}$ Protein (25-35)-induced Neurotoxicity in Cultured Rat Cortical Neurons (Amyloid ${\beta}$ 2 Protein (25-35) 유도 배양신경세포 독성에 대한 목단피의 억제효과)

  • Kim, Joo-Youn;Ju, Hyun-Soo;Ban, Ju-Yeon;Song, Kyung-Sik;Seong, Yeon-Hee
    • Korean Journal of Medicinal Crop Science
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    • v.16 no.6
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    • pp.409-415
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    • 2008
  • Moutan cortex, the root bark of Paeonia suffruticosa Andrews (Paeoniaceae), has pharmacological effects such as anti-inflammatory, antiallergic, analgesic and antioxidant activities. We investigated a methanol extract of Moutan cortex for neuroprotective effects on neurotoxicity induced by amyloid ${\beta}$ protein ($A{\beta}$) (25-35) in cultured rat cortical neurons. Exposure of cultured cortical neurons to $10\;{\mu}M\;A{\beta}$ (25-35) for 24 h induced neuronal apoptotic death. Moutan cortex inhibited $10\;{\mu}M\;A{\beta}$ (25-35)-induced neuronal cell death at 30 and $50\;{\mu}g/m{\ell}$, which was measured by a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay and Hoechst 33342 staining. Moutan cortex inhibited $10\;{\mu}M\;A{\beta}$ (25-35)-induced elevation of intracellular calcium concentration ($[Ca^{2+}]_i$), and generation of reactive oxygen species (ROS) which were measured by fluorescent dyes. Moutan cortex also inhibited glutamate release into medium induced by $10\;{\mu}M\;A{\beta}$ (25-35), which was measured by HPLC. These results suggest that Moutan cortex prevents $A{\beta}$ (25-35)-induced neuronal cell damage by interfering with the increase of $[Ca^{2+}]_i$, and then inhibiting glutamate release and ROS generation. Moutan cortex may have a therapeutic role in preventing the progression of Alzheimer's disease.

Studies on Milk Protein by Zone Electrophoresis (대전기영동법(帶電氣泳動法)에 의한 유청단백질(乳淸蛋白質)에 관(關)한 연구(硏究))

  • Cho, Duk-Bong;Nam, Hyun-Keun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.5 no.1
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    • pp.25-30
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    • 1976
  • In this investigation, the physico-chemical properties of Korean cattles milk and Holstein cows milk are studied. The results of the milk -protein of F-1, F-2, and F-4 hybrid are interested in this studied. 1. The physico-chemical properties of Korean cattle and Holstein cows milk is summarized at Table 1, Table 2. 2. The electrophoretic milk protein fraction of Korean cattle is identified : Lactoalbumin 1.74mg%. Alpha-1 globulin 1.19mg%, Alpha-2 globulin 0.68mg%, Beta-globulin 2.57mg%, Gamma-globulin 0.93mg%. 3. The electrophoretic milk protein fraction of Holstein cow : Lactoalbumin 1.23mg%, Alpha-1 globulin 0.93mg%, Alpha-2 globulin 0.86mg%, Bata-globulin 2.56mg%, Gamma-globulin 1.74mg% have been identified. 4. The electrophoretic milk protein fraction of hybrid F-1, F-2, and F-4 between Korean cattle(female) and Holstein(male) is obtained the following : F-1 hybrid : Lactoalbumin 1.38mg%, Alpha-1 globulin 1.58mg%, Alpa-2 globulin 2.46mg%, Beta-globulin 1.39mg%. F-2 hybrid : Lactoalbumin 1.57mg%, Alpha-1 globulin 1.74mg%, Alpha-2 globulin 2.19mg%, Beta-globulin 1.74mg%. F-4 hybrid : Lactoalbumin 2.46mg%, Alpha-1 globulin 1. 24mg%, Alpha-2 globulin 1.57㎎%, Beta-globulin 1.74mg%.

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Expressions of Tumor-Related Proteins and $TGF-{\beta}1$ in Colon Cancer

  • Kim, Tai-Jeon;Kim, Tae-Geun
    • Biomedical Science Letters
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    • v.13 no.3
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    • pp.213-221
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    • 2007
  • This study was designed to investigate the correlation between the expression rate of p53 and p21 proteins, c-erbB-2 oncoprotein and $TGF-{\beta}1$ and tumor prognostic factors in colon cancer including the tumor size, histological differentiation and Dukes' stage. The expression rate of p53 protein was 11.4% (4 cases) at well differentiation, 48.6% (17 cases) at moderately differentiation, and 17.1% (6 cases) at poorly differentiation. In other words, the poorer differentiation, the higher the expression rate of p53 protein (P<0.05). The expression rate of p21 protein was 17.1% (6 cases) at well differentiation, 40.0% (14 cases) at moderately differentiation, and 8.6% (3 cases) at poorly differentiation, indicating that, as the histological malignant degeneration progressed, the expression rate of p21 protein decreased distinctively (P<0.05). However, the correlation of the above mentioned proteins with tumor size and Dukes' stage was not recognized. The expression rate of c-cerbB-2 oncoprotein was 11.4% (4 cases) at well differentiation, 54.3% (19 cases) at moderately differentiation, and 17.1% (6 cases) at poorly differentiation, indicating that the poorer differentiation, the higher expression rate of c-erbB-2 oncoprotein (P<0.05). The expression rate of $TGF-{\beta}1$ was 17.1% (6 cases) at well differentiation, 48.6% (17 cases) at moderately differentiation, and 11.4% (4 cases) at poorly differentiation. As Dukes' stage progressed, the expression rate of $TGF-{\beta}1$ was 8.6% (3 cases) in stage A, 20.0% (7 cases) in stage B, 37.1 % (13 cases) in stage C, and 11.4% (4 cases) in stage D. There was a difference in expression rates between Dukes' stages (P<0.05). In 10 cases, p53 protein was positive while p21 protein was negative, and in 6 cases, p53 protein was negative whereas p21 was positive (P<0.05). Therefore, a statistically significant inverse correlation between the expression rate of p53 protein and that of p21 protein was observed. In conclusion, since there was a signigicant correlation between histological differentiation of colon cancer and the expressions of p53 and p21 proteins and c-erbB-2 oncoprotein, and between Dukes' stage and the expression of $TGF-{\beta}1$, it was conformed that the overexpression of p53 and p21 proteins, c-erbB-2 oncoprotein, and $TGF-{\beta}1$ is closely associated with the occurrence of colon cancer and its progress. Accordingly, this study may be greatly beneficial to the presumption of diagnosis, treatment and prognosis of colon cancer patients.

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Production and Characterization of a Monoclonal Antibody against Human ${\beta}_2$-adrenergic receptor

  • Kang, Suk-Jo;Shin, Chan-Young;Song, Mi-Ryoung;Lee, Chung-Jae;Cheong, Jae-Hoon;Lee, Sang-Bong;Ko, Kwang-Ho
    • Biomolecules & Therapeutics
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    • v.5 no.4
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    • pp.344-350
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    • 1997
  • The purpose of the present study was to produce and characterize a monoclonal antibody against human ${\beta}_2$-adrenergic receptor. Male BALB/c mice were immunized with glutathione S-transferase (GST) fusion protein of the C-terminal portion of the human ${\beta}_2$-adrenergic receptor which was expressed in E.Coli. The immunized splenocytes were fused with myeloma SP2/0-Agl4 cells. The resulting hybridomas were screened for the production of a monoclonal antibody which can recognize human ${\beta}_2$-adrenergic receptor, and then subcloned by limiting dilution. The resulting monoclonal antibody was named as mAb$\beta$CO2. The mono-clonal antibody $\beta$CO2 was determined as IgM subtype and then purified by anti-mouse IgM-agarose affinity chromatography. The results of ELISA, Western blot, and immunocytochemistry showed that mAb$\beta$CO2 recognized human ${\beta}_2$-adrenergic receptor in the ${\beta}_2$-adrenergic receptor-GST fusion protein and human spider-moid carcinoma cell line A431 with highly specific immunoreactivity, The monoclonal antibody $\beta$CO2 may provide useful tools for the study of the $\beta$-adrenergic receptor of human and other species including rats.

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Historical Overview of the Effect of β-Adrenergic Agonists on Beef Cattle Production

  • Johnson, Bradley J.;Smith, Stephen B.;Chung, Ki Yong
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.5
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    • pp.757-766
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    • 2014
  • Postnatal muscle hypertrophy of beef cattle is the result of enhanced myofibrillar protein synthesis and reduced protein turnover. Skeletal muscle hypertrophy has been studied in cattle fed ${\beta}$-adrenergic agonists (${\beta}$-AA), which are receptor-mediated enhancers of protein synthesis and inhibitors of protein degradation. Feeding ${\beta}$-AA to beef cattle increases longissimus muscle cross-sectional area 6% to 40% compared to non-treated cattle. The ${\beta}$-AA have been reported to improve live animal performance, including average daily gain, feed efficiency, hot carcass weight, and dressing percentage. Treatment with ${\beta}$-AA increased mRNA concentration of the ${\beta}_2$ or ${\beta}_1$-adrenergic receptor and myosin heavy chain IIX in bovine skeletal muscle tissue. This review will examine the effects of skeletal muscle and adipose development with ${\beta}$-AA, and will interpret how the use of ${\beta}$-AA affects performance, body composition, and growth in beef cattle.

Mercury induced the Accumulation of Amyloid Beta (Aβ) in PC12 Cells: The Role of Production and Degradation of Aβ

  • Song, Ji-Won;Choi, Byung-Sun
    • Toxicological Research
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    • v.29 no.4
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    • pp.235-240
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    • 2013
  • Extracellular accumulation of amyloid beta protein ($A{\beta}$) plays a central role in Alzheimer's disease (AD). Some metals, such as copper, lead, and aluminum can affect the $A{\beta}$ accumulation in the brain. However, the effect of mercury on $A{\beta}$ accumulation in the brain is not clear. Thus, this study was proposed to estimate whether mercury concentration affects $A{\beta}$ accumulation in PC12 cells. We treated 10, 100, and 1000 nM $HgCl_2$ (Hg) or $CH_3HgCl_2$ (MeHg) for 48 hr in PC12 cells. After treatment, $A{\beta}_{40}$ in culture medium increased in a dose- and time-dependent manner. Hg and MeHg increased amyloid precursor protein (APP), which is related to $A{\beta}$ production. Neprilysin (NEP) levels in PC12 cells were decreased by Hg and MeHg treatment. These results suggested that Hg induced $A{\beta}$ accumulation through APP overproduction and reduction of NEP.

Cytokine Production in Cholangiocarcinoma Cells in Response to Clonorchis sinensis Excretory-Secretory Products and Their Putative Protein Components

  • Pak, Jhang Ho;Lee, Ji-Yun;Jeon, Bo Young;Dai, Fuhong;Yoo, Won Gi;Hong, Sung-Jong
    • Parasites, Hosts and Diseases
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    • v.57 no.4
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    • pp.379-387
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    • 2019
  • Clonorchis sinensis is a carcinogenic human liver fluke that promotes hepatic inflammatory environments via direct contact or through their excretory-secretory products (ESPs), subsequently leading to cholangitis, periductal fibrosis, liver cirrhosis, and even cholangiocarcinoma (CCA). This study was conducted to examine the host inflammatory responses to C. sinensis ESPs and their putative protein components selected from C. sinensis expressed sequenced tag (EST) pool databases, including $TGF-{\beta}$ receptor interacting protein 1(CsTRIP1), legumain (CsLeg), and growth factor binding protein 2 (CsGrb2). Treatment of CCA cells (HuCCT1) with the ESPs or bacterial recombinant C. sinensis proteins differentially promoted the secretion of proinflammatory cytokines ($IL-1{\beta}$, IL-6, and $TNF-{\alpha}$) as well as anti-inflammatory cytokines (IL-10, $TGF-{\beta}1$, and $TGF-{\beta}2$) in a time-dependent manner. In particular, recombinant C. sinensis protein treatment resulted in increase (at maximum) of ~7-fold in $TGF-{\beta}1$, ~30-fold in $TGF-{\beta}2$, and ~3-fold in $TNF-{\alpha}$ compared with the increase produced by ESPs, indicating that CsTrip1, CsLeg, and CsGrb2 function as strong inducers for secretion of these cytokines in host cells. These results suggest that C. sinensis ESPs contribute to the immunopathological response in host cells, leading to clonorchiasis-associated hepatobiliary abnormalities of greater severity.

The β Subunit of Heterotrimeric G Protein Interacts Directly with Kinesin Heavy Chains, Kinesin-I (Kinesin-I의 kinesin heavy chains과 직접 결합하는 heterotrimeric G protein의 β subunit의 규명)

  • Seog, Dae-Hyun
    • Journal of Life Science
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    • v.20 no.8
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    • pp.1166-1172
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    • 2010
  • Kinesin-I exists as a tetramer of two heavy chains (KHCs, also called KIF5s), which contain the amino (N)-terminal motor domain and carboxyl (C)-terminal domain, as well as two light chains (KLCs), which bind to the KIF5s (KIF5A, KIF5B and KIF5C) stalk region. To identify the interaction proteins for KIF5A, yeast two-hybrid screening was performed and a specific interaction with the ${\beta}$ subunit of heterotrimeric G proteins ($G{\beta}$) was found. $G{\beta}$ bound to the amino acid residues between 808 and 935 of KIF5A and to other KIF5 members in the yeast two-hybrid assay. The WD40 repeat motif of $G{\beta}$ was essential for interaction with KIF5A. In addition, these proteins showed specific interactions in the glutathione S-transferase (GST) pull-down assay. An antibody to KIF5s specifically co-immunoprecipitated KIF5s associated with heterotrimeric G proteins from mouse brain extracts. These results suggest that kinesin-I motor protein transports heteroterimeric G protein attachment vesicles along microtubules in the cell.

Incremental Expression of 14-3-3 Protein Beta/Alpha in Urine Correlates with Advanced Stage and Poor Survival in Patients with Clear Cell Renal Cell Carcinoma

  • Kaneko, Shiho;Matsumoto, Kazumasa;Minamida, Satoru;Hirayama, Takahiro;Fujita, Tetsuo;Kodera, Yoshio;Iwamura, Masatsugu
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.3
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    • pp.1399-1404
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    • 2016
  • We investigated the urinary levels of 14-3-3 protein beta/alpha to evaluate their diagnostic significance with regard to clear cell renal cell carcinoma (ccRCC) and angiomyolipoma (AML). Urine samples from 91 patients with ccRCC, 16 patients with AML and 24 healthy volunteers were assessed. We used an enzyme-linked immunosorbent assay (ELISA) to quantify 14-3-3 protein beta/alpha levels in urine. Values were higher in patients with ccRCC than in those with AML and in healthy volunteers. High levels were associated with pathologic stage, lymph node status, distant metastasis and poor survival. Urinary levels of 14-3-3 protein beta/alpha were significantly increased in patients with small-sized carcinoma, irrespective of being less than 4.0 cm and 2.0 cm, compared with levels in patients with AML. This study is the first to report that increased expression of 14-3-3 protein beta/alpha in urine is associated with advanced stage and poor survival in patients with ccRCC. In addition, urinary 14-3-3 protein beta/alpha may differentiate AML from RCC, even when small sized. These results suggest that examination of urinary 14-3-3 protein beta/alpha could serve as a diagnostic and prognostic marker in patients with ccRCC.