• Title/Summary/Keyword: S-100 Protein

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Characterization and Cloning of the Gene Encoding Autoregulator Receptor Protein from Streptomyces longwoodensis (Streptomyces longwoodensis로부터 Autoregulator Receptor Protein 유전자의 클로닝 및 특성)

  • Yeo Soo-Hwan;Lee Sung-Bong;Kim Hyun-Soo
    • Microbiology and Biotechnology Letters
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    • v.33 no.2
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    • pp.96-105
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    • 2005
  • For screening of autoregulator receptor gene from Streptomyces longwoodensis, PCR was performed with primers of receptor gene designed on the basis of amino acid sequences of autoregulator receptor proteins with known function. PCR products were subcloned into the BamHIsite of pUC19 and transformed into the E. coli $DH5{\alpha}$. The isolated plasmid from transformant contained the fragment of 100 bp, which was detected on $2\%$ gel after BamHI treatment. The insert, 100 bp PCR product, was confirmed as the expected internal segment of gene encoding autoregulator receptor protein by sequencing. Southern and colony hybridizations with the 100 bp fragment as a probe allowed to select a genomic clone of S. longwoodensis, pSLT harboring a 4.4 kb SphI fragment. Nucleotide sequencing analyses revealed a 651 bp open reading frame(ORF) were isolated protein showing moderate homology ($35{\sim}46\%$) with the ${\Gamma}$-butyrolactone autoregulator receptors from Streptomyces sp., and this ORF was named sltR The sltR/pET-17b plasmid was constructed to overexpress the recombinant SltR protein (rSltR) in E. coli BL21 (DE3)/pLysS, and the rSltR protein was purified to homogeneity by DEAE-Sephacel column chromatography, and DEAE-5PW chromatography (HPLC). The molecular mass of the purified rSltR protein was 55 kDa by HPLC gel-filtration chromatography and 28 kDa by SDS-PAGE, indicating that the rSltR protein is present as a dimer. A binding assay with tritium-labeled autoregulators revealed that the rSltR has clear binding activity with a A-factor type autoregulator as the most effective ligand.

The Relationship between Milk Protein Phenotypes and Lactation Traits in Brown Swiss and Canadienne

  • Kim, S.;Ng-Kwai-Hang, K.F.;Hayes, J.F.
    • Asian-Australasian Journal of Animal Sciences
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    • v.11 no.3
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    • pp.311-317
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    • 1998
  • A total of 1033 Brown Swiss and 610 Canadienne cows were phenotyped for the genetic variants ${\alpha}_{s1}$-casein, ${\beta}$-casein, ${\kappa}$-casein, ${\beta}$-lactoglobulin and ${\alpha}$-lactalbumin. In Brown Swiss, frequency distributions were: 97.3% B and 2.7% C variant of ${\alpha}_{s1}$-casein; 31.6% $A^1$, 51.8% $A^2$, 0.5% $A^3$ and 16.1% B variant of ${\beta}$-casein; 70.4% A, 29.3% B, and 0.3% C variant of ${\kappa}$-casein; 41.7% A and 58.3% B variant of ${\beta}$-lactoglobulin; and 100% B variant of ${\alpha}$-lactalbumin. Corresponding frequencies in Canadienne for those five milk proteins were: 98.6 and 1.4%;58.5, 33.5, 0.08 and 7.9%; 78.8, 21.1 and 0.1%, 42.4 and 57.6%; and 100%. Analysis of variance by least squares showed possible association between milk protein phenotypes and some lactational production traits. There were no significant association of phenotypes of ${\alpha}_{s1}$-casein, ${\beta}$-casein and ${\beta}$-lactoglobulin with milk yield, fat yield, protein yield, fat percentage and protein percentage in both breeds during the three lactations. In the Brown Swiss, ${\kappa}$-casein phenotype was associated with 305-day fat yield and protein yield during the first lactation. ${\kappa}$-Casein AB was associated with higher milk, fat and protein yield during the second lactation. During the third lactation, ${\beta}$-lactoglobulin AA in Canadienne cows was associated with higher protein content in the milk (3.70%) when compared to phenotypes AB (3.54%) and BB (3.64%).

Characteristics of the protein and antioxidant contents in soybeans cultured with Phellinus linteus HN00K9 (목질진흙버섯 (Phellinus linteus) HN00K9 균사체 배양 대두의 단백질함량 및 항산화활성 특성)

  • Ja-Yoon Kim;Ye-Lin Baek;Sang-Ho Lee;Hee-Wan Kang
    • Journal of Mushroom
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    • v.21 no.3
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    • pp.126-131
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    • 2023
  • In this study, the protein content and functional changes in soybeans cultured with Phellinus linteus HN00K9 were analyzed. P. linteus HN00K9 was cultured on soybeans. The crude protein content in soybeans cultured with HN00K9 (PMS) was 41.99%, which was higher than that in soybeans not cultured with the mushroom (UCS). The total free amino acid content in PMS increased to 39,963 mg/100 g, which was higher than that in UCS (36,817 mg/100 g). In particular, in PMS, glutamic acid accounted for 18.5% of the total amino acids at 7,413 mg/100 g. The total polyphenol content in PMS was 2.66 mg GAE/g, which was more than 45% higher than the amount in UCS (1.45 mg GAE/g). Additionally, PMS showed a DPPH radical scavenging activity of 33.3%, which was 3 times higher than that exhibited by UCS (11.5%), reflecting its high antioxidant content. Therefore, the PMS in this study has potential for use as a functional food material.

The Fundamental Studies of Heat Treatment Class in Domestic Infant Formula (국산 조제분유의 열처리 등급에 관한 기초 연구)

  • 박영희
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.3
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    • pp.495-500
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    • 1998
  • To compare the extent of heat treatment in domestic infant formula, pH,titratable acidity, undenatured whey protein contents, HMF contents and protein-reducing substances of three commercial products (A, B, C) were measured. The pH of B products was lowest and the titratable acidity of B product was highest. The contents of undenatured whey protein per 100ml serum were 0∼30mg(A products), 90∼130mg(B products)and 80∼90mg(C products), respectively. Distinct differences of undenatured whey protein contents according to the manufacturer and infat's stage in age could be observed. The HMF contents of tested products showed 10.9∼21.5umol/L and B-2 product(B products for the second stage of 5∼9 month) was the highest among tested products. The protein-reducing substances showed 4.46∼9.50mg K4Fe(CN)6/100ml serum nd B-2 product was the highest among tested products.

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Production of Pectolytic Enzymes and Change of Pectic Substances from Apple Fruits Infected with Botryosphaeria dothidea (사과 겹무늬썩음병균(Botryosphaeria dothidea)에 의해 부패된 사과 과실에서 Pectin질 분해효소의 생산과 Pectin질의 변화)

  • Park, Seok-Hee;Lee, Chang-Un
    • The Korean Journal of Mycology
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    • v.21 no.2
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    • pp.106-111
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    • 1993
  • Pectolytic enzymes were extracted in apple fruits rotted by Botryosphaeria dothidea, and their activities and change of pectic substances were investigated. Exo-polygalacturonase(exo-PG), exo-polymethylgalacturonase(exe-PMG), polygalacturonate-trans-eliminase(PGTE) and pectin-methyl-trans-eliminase(PMTE) were produced by the pathogen. Activities of exo-PG and exe-PMG extracted from rotten apple fruits were high to 21.15 and 24.65 units/mg protein in specific activity at seven days after inoculation, respectively. Activities of PGTE and PMTE showed 5.60 and 7.90 units/mg protein, respectively, but they were lower than those of the exo-type enzymes. Water-soluble and versene-soluble pectins were 11.50 mg/100 mg-AIS and 7.31 mg/100 mg-AIS at 14 days after inoculation, namely, they were increased by 4.23 and 2.16 mg/100 mg-AIS over those of sound apples, respectively. Total soluble pectic substances of rotten apple were 72.4% of total pectic substances and it was higher by 24.8% than sound apple. Insoluble pectic substance was notably decreased from 15.32 to 7.16 mg/100 mg-AIS according to progress of decay while total pectic substances were not changed remarkably.

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Histological and immunohistochemical studies on the gizzard and pylorus regions of the chicken embryos (닭 태자의 근육위와 유문부에 대한 조직학적 및 면역조직화학적 연구)

  • Lee, Jae-hyun;Ku, Sae-kwang;Lee, Hyeung-sik
    • Korean Journal of Veterinary Research
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    • v.38 no.4
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    • pp.686-695
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    • 1998
  • Histological changes, distributions and relative frequencies of bovine Sp-1/chromogranin (bCG)-, serotonin-, gastrin-, cholecystokinin-8(CCK-8)-, somatostatin-, S-100 protein-, polypeptide YY(PYY)- and glucagon-immunoreactive cells were investigated in the gizzard and pylorus of the chicken embryos from 10 days of incubation to hatching. Histologically, the pseudostratified columnar epithelium were observed from 10 days of incubation to 15 days of incubation, thereafter these epithelium were differentiated to simple columnar epithelium, gastric gland and/or mucosal gland. In the gizzard, bCG-immunoreactive cells were observed from 19 days of incubation and S-100 protein-immunoreactive cells were detected from 15 days of incubation to 18 days of incubation. No serotonin-, gastrin-, CCK-8-, somatostatin-, PYY- and glucagon-immunoreactive cells were found in this region. In the pylorus, bCG-, gastrin- and somatostatin-immunoreactive cells were observed from 16 days of incubation respectively, thereafter these cells were increased with ages. CCK-8-immunoreactive cells were detected on hatching and S-100 protein-immunoreactive cells were detected from 16 days of incubation to 18 days of incubation. No serotonin-, PYY- and glucagon-immunoreactive cells were observed in this region.

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Stimulation of Ribosome Activity of Zea mays by Polyamine (Polyamine에 의한 옥수수 Ribosome의 활성 촉진)

  • 김기남
    • Journal of Plant Biology
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    • v.36 no.1
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    • pp.83-90
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    • 1993
  • As a part of the study on the relation between exogenous polyamines and various components necessary for protein biosynthesis in the germinating maize seeds, the effects of the polyamines on protein biosynthesis and irbosome activity were investigated. The protein biosynthesis activity by S-30 containing all components necessary for protein biosynthesis was increased by exogenous polyamines, spermidine, spermine and putrescine. As the concentration of polyamine treated was increased, the optimal Mg2+ concentration of in vitro poly U-dependent protein synthesis system was gradually reduced. However, the optimal Mg2+ concentration of poly U-dependent system containing optimal polyamine was 10 mM regardless of the sort of polyamine. It could be infered that polyamines play an important part in protein biosynthesis in the higher plant, and that the role of polyamines take partially the place of Mg2+ action. The activities of ribosome and S-100 in protein biosynthesis were increased by 46.7% and 17.7% with spermidine, and by 44.1% and 16.2% with spermine, and by 29.1% and 19.3% with putrescine. It could be concluded that the increase of protein biosynthesis by polyamines in mainly owing to the ribosome activation.

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A Growth-Stimulating Protein in Cow's Milk

  • Ki, Eun-Kyung;Park, Yong-Bok;Choi, Myung-Sook
    • Journal of the East Asian Society of Dietary Life
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    • v.4 no.3
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    • pp.59-65
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    • 1994
  • Growth-stimulating effects of cow's milk was examined using Vero cell cultre. Medium containing whole cow's milk stimulated cell growth about the same degree as that containing fetal bovine serum. The growth-stimulating factor in cow's milk was purified using hydrophobic (phenyl-sepharose) and gel filtration (Sephadex G-100) column chromatographies. It appeared that the factor is a highly hydrophobic protein, since the major growth-stimulating activity was found in the fractions eluted with 50% ethylene glycol from the phenyl-sepharose column during the purification. The purified factor showed a single band on the polyacrylamide gel electrophoresis in the presence of 1% (w/v) SDS. The factor has been found to have a relatively high molecular weight in the range of about Mr=100,000-150,000. In the presence of the purified factor (5%, w/v) in the culture medium, the incorporation of [3H]-thymidine into the cells was increased approximately 2,400-fold over that in the presence of 5% (w/v) fetal bovine serum. It seems that the growth-stimulating factor purified in this study is one of the major growth factors in the cow's milk.

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S100ß, Matrix Metalloproteinase-9, D-dimer, and Heat Shock Protein 70 Are Serologic Biomarkers of Acute Cerebral Infarction in a Mouse Model of Transient MCA Occlusion

  • Choi, Jong-Il;Ha, Sung-Kon;Lim, Dong-Jun;Kim, Sang-Dae;Kim, Se-Hoon
    • Journal of Korean Neurosurgical Society
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    • v.61 no.5
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    • pp.548-558
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    • 2018
  • Objective : Diagnosing acute cerebral infarction is crucial in determining prognosis of stroke patients. Although many serologic tests for prompt diagnosis are available, the clinical application of serologic tests is currently limited. We investigated whether $S100{\beta}$, matrix metalloproteinase-9 (MMP-9), D-dimer, and heat shock protein 70 (HSP70) can be used as biomarkers for acute cerebral infarction. Methods : Focal cerebral ischemia was induced using the modified intraluminal filament technique. Mice were randomly assigned to 30-minute occlusion (n=10), 60-minute occlusion (n=10), or sham (n=5) groups. Four hours later, neurological deficits were evaluated and blood samples were obtained. Infarction volumes were calculated and plasma $S100{\beta}$, MMP-9, D-dimer, and HSP70 levels were measured using enzyme-linked immunosorbent assay. Results : The average infarction volume was $12.32{\pm}2.31mm^3$ and $46.9{\pm}7.43mm^3$ in the 30- and 60-minute groups, respectively. The mean neurological score in the two ischemic groups was $1.6{\pm}0.55$ and $3.2{\pm}0.70$, respectively. $S100{\beta}$, MMP-9, and HSP70 expressions significantly increased after 4 hours of ischemia (p=0.001). Furthermore, $S100{\beta}$ and MMP-9 expressions correlated with infarction volumes (p<0.001) and neurological deficits (p<0.001). There was no significant difference in D-dimer expression between groups (p=0.843). The area under the receiver operating characteristic curve (AUC) showed high sensitivity and specificity for MMP-9, HSP70 (AUC=1), and $S100{\beta}$ (AUC=0.98). Conclusion : $S100{\beta}$, MMP-9, and HSP70 can complement current diagnostic tools to assess cerebral infarction, suggesting their use as potential biomarkers for acute cerebral infarction.