• Title/Summary/Keyword: protein A

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Expression and Characterization of Uropathogenic Escherichia coli Adhesin Protein Linked to Cholera Toxin A2B Subunits in Escherichia coli TB1

  • Lee, Yong-Hwa;Ryu, Dong-Kyun;Kim, Byung-Oh;Pyo, Suhk-Neung
    • Journal of Microbiology and Biotechnology
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    • v.13 no.4
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    • pp.552-559
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    • 2003
  • The FimH subunit of type 1-fimbriated Escherichiu coli (E. coli) has been determined as a major cause for urinary tract infections. Thus, to produce a possible vaccine antigen against urinary tract infections, the fimIH gene was genetically coupled to the ctxa2b gene and cloned into a pMAL-p2E expression vector. The chimeric construction of pMALfimH/ctxa2b was then transformed into E. coli K-12 TB1 and its nucleotide sequence was verified. A fusion protein, based on fusing adhesin to the cholera toxin subunit A2B (CTXA2B), was induced with 0.01 mM isopropyl-${\beta}-D-thiogalactoside$ (IPTG) for 4 h at $37^{\circ}C$ to yield a soluble fusion protein. The fusion protein was then purified by affinity chromatography. The expressed fusion protein was confirmed by SDS-PAGE and Western blotting using antibodies to the maltose binding protein (MBP) or the cholera toxin subunit B (CTXB), plus the N-terminal amino acid sequence was also analyzed. The orderly-assembled fusion protein was confirmed by a modified $G_{Ml}-ganglioside$ ELISA, using antibodies to adhesin. The results indicated that the purified fusion protein was an adhesin/CTXA2B protein containing E. coli adhesin and the $G_{Ml}-ganglioside$ binding activity of CTXB. Accordingly, this adhesin/CTXA2B protein may be a potential antigen for oral immunization against uropathogenic E. coli.

A Fusion Tag to Fold on: The S-Layer Protein SgsE Confers Improved Folding Kinetics to Translationally Fused Enhanced Green Fluorescent Protein

  • Ristl, Robin;Kainz, Birgit;Stadlmayr, Gerhard;Schuster, Heinrich;Pum, Dietmar;Messner, Paul;Obinger, Christian;Schaffer, Christina
    • Journal of Microbiology and Biotechnology
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    • v.22 no.9
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    • pp.1271-1278
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    • 2012
  • Genetic fusion of two proteins frequently induces beneficial effects to the proteins, such as increased solubility, besides the combination of two protein functions. Here, we study the effects of the bacterial surface layer protein SgsE from Geobacillus stearothermophilus NRS 2004/3a on the folding of a C-terminally fused enhanced green fluorescent protein (EGFP) moiety. Although GFPs are generally unable to adopt a functional confirmation in the bacterial periplasm of Escherichia coli cells, we observed periplasmic fluorescence from a chimera of a 150-amino-acid N-terminal truncation of SgsE and EGFP. Based on this finding, unfolding and refolding kinetics of different S-layer-EGFP chimeras, a maltose binding protein-EGFP chimera, and sole EGFP were monitored using green fluorescence as indicator for the folded protein state. Calculated apparent rate constants for unfolding and refolding indicated different folding pathways for EGFP depending on the fusion partner used, and a clearly stabilizing effect was observed for the SgsE_C fusion moiety. Thermal stability, as determined by differential scanning calorimetry, and unfolding equilibria were found to be independent of the fused partner. We conclude that the stabilizing effect SgsE_C exerts on EGFP is due to a reduction of degrees of freedom for folding of EGFP in the fused state.

Synthesis of Nitrogen Doped Protein Based Carbon as Pt Catalysts Supports for Oxygen Reduction Reaction (산화환원반응용 백금 촉매 지지체를 위한 질소 도핑된 단백질계 탄소의 제조)

  • Lee, Young-geun;An, Geon-hyeong;Ahn, Hyo-Jin
    • Korean Journal of Materials Research
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    • v.28 no.3
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    • pp.182-188
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    • 2018
  • Nitrogen (N)-doped protein-based carbon as platinum (Pt) catalyst supports from tofu for oxygen reduction reactions are synthesized using a carbonization and reduction method. We successfully prepare 5 wt% Pt@N-doped protein-based carbon, 10 wt% Pt@N-doped protein-based carbon, and 20 wt% Pt@N-doped protein-based carbon. The morphology and structure of the samples are characterized by field emission scanning electron microscopy and transmission electron micro scopy, and crystllinities and chemical bonding are identified using X-ray diffraction and X-ray photoelectron spectroscopy. The oxygen reduction reaction are measured using a linear sweep voltammogram and cyclic voltammetry. Among the samples, 10 wt% Pt@N-doped protein-based carbon exhibits exellent electrochemical performance with a high onset potential of 0.62 V, a high $E_{1/2}$ of 0.55 V, and a low ${\Delta}E_{1/2}=0.32mV$. Specifically, as compared to the commercial Pt/C, the 10 wt% Pt@N-doped protein-based carbon had a similar oxygen reduction reaction perfomance and improved electrochemical stability.

Transcription Factor PU.1 Inhibits Aspergillus fumigatus Infection via Surfactant Protein-D

  • Kim, Sung-Su
    • Biomedical Science Letters
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    • v.24 no.3
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    • pp.175-182
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    • 2018
  • Aspergillosis is a life-threatening disease in individuals with compromised immune systems. Fungal invasion is a highly critical process during host cellular infection. Several papers have reported that transcription factors are responsible for the infection process. To investigate what transcription factors are involved in the process in an effort to inhibit fungal infection into cells, I checked the surfactant protein family and PU.1 transcription factor levels in A549 cells infected with A. fumigatus conidia. PU.1 and surfactant protein-D levels were reduced in cells infected with fungal conidia. I then observed an increase in surfactant protein-D on PU.1-overexpressed cells. Infection of A. fumigatus conidia was decreased in PU.1-overexpressed cells, whereas the suppression of PU.1 did not lead to any changes in cases of A. fumigatus conidia infection. These results indicate that PU.1 inhibits the infection of A. fumigatus conidia via the expression of surfactant protein-D, suggesting that PU.1 is a key transcription factor for protection against A. fumigatus invasion.

An Effect of Bromobenzene Treatment on the Liver Damage of Rats Previously Fed Low or High Protein Diet (단백식이 조건을 달리하여 성장시킨 흰쥐에 Bromobenzene 투여가 간손상에 미치는 영향)

  • 신중규;채순님;윤종국
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.23 no.6
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    • pp.894-898
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    • 1994
  • To evaluate an effect of dietary protein on the liver damage, the bromobenzene was intraperitoneally injected to the rats fed a low or high protein diet and then the liver weight per body weight and serum levels of alanine aminotransferase (ALT) activities were determined to demonstrate the differences in liver damage between the groups fed low or high protein diet. Hepatic aniline hydroxylase (AH), glutthione (GSH) content and glutathione s-transferase(GST) activity were also determined to clarify causes of liver damage between the two groups. Increases of liver weight per body weight and serum ALT activities were higher in brombenzene treated rats fed low protein diet than those fed high protein diet. The increasing rate of hepatic AH activity was higher in bromobenzne-treated rats fed low protein diet than that in those fed high protein diet. Furthermore , hepatic glutathione contents and GST activities in bromobenzene-treated rats were higher in rats fed high protein diet than those fed low protein diet. In case of control group, the heaptic glutathione content and GST activity were also higher in rats fed high protein diet than those fed low protein diet.

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Dietary Reference Intakes for Protein: Protein Requirement and Estimation Method, AMDR (Amount of Macronutrient Distribution Range), for Protein (단백질 섭취기준: 단백질 필요량과 추정 방법 및 단백질에너지 적정비율)

  • Chang, Soon-Ok
    • Journal of Nutrition and Health
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    • v.44 no.4
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    • pp.338-343
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    • 2011
  • This study assessed the current EAR, RDA, and AMDR for protein, which were set in 2005 and revised in 2010 as the DRIs for Koreans. A classical approach to establish the EAR for protein has been the nitrogen balance method. This method has practical limitations and problems in statistical analysis by giving over estimations of nitrogen balance. Thus, the present EAR for protein might be lower than the true requirement. Recent reevaluations of nitrogen balance studies by bilinear regression analysis and the IAAO method have indicated that the EAR of 0.66 g/kg bw/d should be increased by 39% to give 0.92 g/kg bw/d. The AMDR for protein in the Korean DRIs was set at 7-10%, which covers almost the entire population's protein intake. Since the 5th percentile of Korean protein intake is close to 10% of energy and due to the beneficial effects of protein beyond the maintenance of nitrogen equilibrium, the lower range of 7% needs to be increased up to 10%. For practical meal arrangement, 15% of energy as protein, which is close to the average protein intake of Koreans, seems to be proper, although the value is almost two times the EAR.

Effects of Dietary Protein on Growth and Lipid Metabolism in Growing Rats (식이단백질조성이 흰쥐의 성장과 지방대사에 미치는 영향)

  • Kim, Yoo-Sook;Kim, Wha-Young
    • Journal of Nutrition and Health
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    • v.15 no.2
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    • pp.119-128
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    • 1982
  • The effect of dietary protein on growth and lipid levels of plasma and liver was studied in weanling male rats fed diets differing protein sources and amino acid balance. Rats were devided into 9 experimental diets which were grouped into 3 categories ; 1) Simple protein category includes gluten-, soy protein isolate-, and casein-containing diet groups, 2) Supplemented category includes casein supplemented with methionine, soy protein isolate supplemented with methionine, and gluten supplemented with lysine and methionine, 3) Mixed protein category includes diet groups containing gluten (2/3), casein (1/3), soy protein isolate (2/3) and casein (1/3), and casein (1/3), soy protein isolate (1/3) and gluten (1/3). The experimental diets composed of 15% protein, 65.8% carbohydrate, 10% fat and 1% cholesterol. The body wt. gain and P.E.R. were greater in rats of supplemented and mixed protein groups than simple protein groups. No statistical differences were found in plasma cholesterol among gluten, soy protein isolate and casein groups. Consumption of diets supplemented with limiting amino acid to gluten or soy protein isolate reduced the plasma cholesterol level by 23.2% and 34.2% respectively. However there was no difference between casein and the supplemented casein groups. The mixed protein groups shows relatively high plasma cholesterol concentration and low liver cholesterol levels. On the other hand gluten group showed low plasma cholesterol and high liver cholesterol levels, which means body cholesterol pool may not have been changed by the dietary protein. Feeding soy protein meal and the supplemented soy protein isolate resulted in lower plasma cholesterol, plasma triglycerides, liver cholesterol and liver triglycerides levels. This hypolipidemic effect is considered to see unique to soy protein isolate. Rats in gluten and the supplemented gluten groups showed lower plasma protein levels and a tendency of fatty liver.

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Regional differences in protein intake and protein sources of Korean older adults and their association with metabolic syndrome using the 2016-2019 Korea National Health and Nutrition Examination Surveys: a cross-sectional study

  • You-Sin Lee;Yoonna Lee
    • Korean Journal of Community Nutrition
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    • v.29 no.3
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    • pp.173-188
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    • 2024
  • Objectives: The study aim was to analyze the regional differences in dietary protein intake and protein sources of Korean older adults and their association with metabolic syndrome. Methods: Study participants were 1,721 older adults aged 65 and over who participated in 2016-2019 Korea National Health and Nutrition Examination Survey. Using 24-hour recall dietary intake data, protein intake and their food sources were examined. The association between protein intake and metabolic syndrome, obesity, and abdominal obesity were analyzed by multiple logistic regression. Results: Total protein and animal protein intakes were higher in urban area (60.0 g, 24.4 g, respectively) than in rural area (54.6 g, 19.6 g, respectively). With increase of protein intake level, animal to total protein proportion was increased in both areas. Total protein and plant protein intake was negatively associated with the risk of obesity, abdominal obesity in both areas. Animal protein intake was negatively associated with the risk of obesity in both areas, and with abdominal obesity only in urban area. In urban area, plant protein intake was also negatively associated with the risks of metabolic syndrome, elevated triglyceride, and reduced high density lipoprotein-cholesterol. In urban area, the risk of metabolic syndrome was decreased when their protein intake was more than 0.91 g/kg and was lowest when their protein intake was more than 1.5 g/kg (P for trend < 0.001). Conclusions: Korean older adults showed inadequate protein intake and those in rural area showed lower animal protein intake than in urban area. The risk of obesity and metabolic syndrome was decreased with the increase of protein intake level. These findings may help develop effective nutrition support strategy for older adults to reduce regional health disparity.

Transcriptomic analysis of 'Campbell Early' and 'Muscat Bailey A' grapevine shoots exposed to freezing cold stress (영하의 저온에 노출된 'Campbell Early'와 'Muscat Bailey A' 포도나무 신초의 전사체 비교)

  • Kim, Seon Ae;Yun, Hae Keun
    • Journal of Plant Biotechnology
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    • v.43 no.2
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    • pp.204-212
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    • 2016
  • To understand the responses of grapevines in response to cold stress causing the limited growth and development, differentially expressed genes (DEGs) were screened through transcriptome analysis of shoots from 2 grapevine cultivars ('Campbell Early' and 'Muscat Baily A') kept at -$2^{\circ}C$ for 4 days. In gene ontology analysis of DEGs from 'Campbell Early', there were 17,424 clones related with biological process, 28,954 with cellular component, and 6,972 with molecular function genes in response to freezing temperature. The major induced genes included dehydrin xero 1, K-box region and MADS-box transcription factor family protein, and MYB domain protein 36, and inhibited genes included light-harvesting chlorophyll B-binding protein 3, FASCICLIN-like arabinoogalactan 9, and pectin methylesterase 61 in 'Campbell Early' grapevines. In gene ontology analysis of DEGs from 'Muscat Baily A', there were 1,157 clones related with biological process, 1,350 with cellular component, and 431 with molecular function gene. The major induced genes of 'Muscat Baily A' included NB-ARC domain-containing disease resistance protein, fatty acid hydrozylase superfamily, and isopentenyltransferase 3, and inhibited genes included binding, IAP-like protein 1, and pentatricopeptide repeat superfamily protein. All major DEGs were shown to be expressed differentially by freezing temperature in real time-PCR analysis. Protein domain analysis using InterPro Scan revealed that ubiquitin-protein ligase was redundant in both tested grapevines. Transcriptome profile of shoots exposed to cold can provide new insights into the molecular basis of tolerance to low-temperature in grapevines, and can be used as resources for development new grapevines tolerant to coldness.

Effects of Low-Calorie Diet Including High Protein-Low Carbohydrate Protein Bar on Weight Loss and Serum Lipid Indicators in Overweight Women according to Dietary Compliance (고단백 저탄수화물식 프로틴바를 이용한 저칼로리 다이어트가 순응 정도에 따라 과체중 여성의 체중감량과 혈청 지질지표에 미치는 영향)

  • Park, Dasom;Lee, Hyun Joo;Son, Sook Mee
    • Korean Journal of Community Nutrition
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    • v.24 no.6
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    • pp.485-496
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    • 2019
  • Objectives: This study was conducted to investigate the effect of a 6-week low-calorie diet (LCD) program including high protein-low carbohydrate protein bar on weight loss, blood pressure, and blood lipid profile in 40 overweight women according to dietary compliance. Methods: Subjects were 62 healthy overweight women (BMI ≥ 23.0 or body fat percentage ≥ 28%), aged 20~59 yrs who were provided a high protein-low carbohydrate protein bar (each 35 g, 154 kcal, protein energy %: 28.6%, carbohydrate energy %: 38.7%) as part of dinner for 6 weeks. Forty subjects who completed the whole diet program were categorized into high compliance (HC) group (days of eating protein bar ≥ 5 weeks) or low compliance (LC) group (days < 5 weeks). Results: Energy intake significantly decreased from 1,867.5 kcal at baseline to 1,137.4 kcal at 6 weeks for the HC group and from 1,971.7 kcal to 1,362.2 kcal for the LC group, respectively. On the other hand, a significant increase in protein energy percentage was observed in each group (HC group: 3.5%, LC group: 2.2%). Both groups showed significant decreases in weight (HC group: 1.8 kg, LC group: 1.1 kg), BMI, fat mass, systolic blood pressure, total cholesterol, and LDL-cholesterol. Reduction of body fat percentage and diastolic blood pressure were only observed in the HC group. Conclusions: The inclusion of a high protein-low carbohydrate protein bar as part of a low-calorie diet for a short period can be effective to achieve weight loss and concomitantly improve blood cholesterol level without serious physiological side effects. More evident results can be achieved by eating a diet with low calorie diet including high protein-low carbohydrate protein bar for more than 5 weeks.