• Title/Summary/Keyword: CAT protein

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Effects of Ginseng Radix on sperm count and CatSper3, 4 proteins expression in Male Mice (인삼(人蔘)이 수컷 생쥐의 정자 수와 CatSper3, 4 단백질 발현에 미치는 영향)

  • Park, Eun-Hwa;Kim, Do-Rim;Park, Seong-Kyu;Chang, Mun-Seog
    • The Korea Journal of Herbology
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    • v.27 no.2
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    • pp.47-51
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    • 2012
  • Objectives : The purpose of this study was to investigate the reproductive effect of Ginseng radix on male mice. Methods : Male C57BL/6J mice were divided into four groups ; normal group (vehicle-treated, n=8), Ginseng radix treatment group (100, 500, 1000 mg/kg, n=8). Ginseng radix extract was treated for 5 weeks. After treatment each group was examined for assessment of sperm count and CatSper protein expression using computer assisted semen analysis (CASA) system and the immunofluorescence. Results : Sperm count of normal and Ginseng radix extract treated group were 287.57 vs. 371.62, 364.83, $343.29{\times}10^6$, respectively. The CatSper3, 4 proteins expression of Ginseng radix treated group were significantly increased than that of normal group. Conclusions : These findings suggest that the Ginseng radix improves male reproductive function by increasing sperm count and CatSper protein expression.

Effect of Dietary Protein and Taurine on Cysteine Catabolism in Cat Liver (식이내의 단백질과 타우린 함량이 Cysteine 대사에 미치는 영향)

  • 박태선
    • Journal of Nutrition and Health
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    • v.29 no.7
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    • pp.729-737
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    • 1996
  • Activieties of hepatic cysteine desulfhydration was assessed in cats fed one of the following diets for 5 weeks : 20% protein, 0% taurine diet(LPOT) ; 20% protein, 0.15% taurine diet (LPNT) ; 60% protein, 0% taurine diet(HPOT) ; and 60% protein, 0.15% taurine diet(HPNT). Cats fed LPOT and HPOT had been maintained on a taurine-free diet for 6 weeks prior to the experiment in order to deplete body taurine. Activities of cysteine desulfhydration were determined by measuring the production of H235S from 35S-cysteine in the presence and absence of $\alpha$-ketoglutarate ($\alpha$-KG) in the incubation medium. The direct pathway via cysteine desulfhydrase appears to account for the major route of cysteine desulfhydration in the cat liver since the values obtained in the absence of $\alpha$-KG were between 81 and 88% of those obtained in the presence of $\alpha$-KG. Mean$\pm$SEM of the hepatic total desulfhydration activities(umol H2S.min-1.kg body wt-1)in cats fed LPOT, LPNT, HPOT and HPNT were 117$\pm$6, 135$\pm$10, 137$\pm$10, and 190$\pm$9, respectively. The capacity of hepatic cysteine desulfhydration (UA/kg body wt) was positively cerrelated not only with the dietary concentration of taurine but also with the concentration of protein.

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Nucleotide Sequence and Inducibility Analysis of Chloramphenicol Acetyltransferase Gene from Staphylococcus aureus R-plasmid pSBK203 (Staphylococcus aureus에서 분리된 R-plasmid pSBK203상의 chloramphenicol acetyltransferase 인자의 염기서열 및 유발성 분석)

  • 권동현;변우현
    • Korean Journal of Microbiology
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    • v.27 no.3
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    • pp.194-200
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    • 1989
  • The nucleotide sequence of inducible chloramphenicol acetyl-transferase(CAT) gene isolated from a small plasmid pSBK203 of Staphylococcus aureus was determined. The base sequence shows that structural gene of pSBK203-CAT encodes a protein of 213 amino acids and has a leader region which encodes a short polypeptide of 9 amino-acids in its upstream. vertical bar /sup 35/S vertical bar-Methionine labelled CAT gene product in minicell showed almost same mobility with pC194-CAT of which molecular weight is 24Kdal on polyacrylamide gel electrophoresis. Predicted amino acid sequence of pSBK203-CAT has revealed a high degree of homology with the CATs of pC194 and pC221 than those of cat-86, Tn9 and proteus mirabilis PM13.

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Effects of chronic caloric restriction on kidney and heart redox status and antioxidant enzyme activities in Wistar rats

  • Dutra, Marcio Ferreira;Bristot, Ivi Juliana;Batassini, Cristiane;Cunha, Nubia Broetto;Vizuete, Adriana Fernanda Kuckartz;Souza, Daniela Fraga De;Moreira, Jose Claudio Fonseca;Goncalves, Carlos-Alberto
    • BMB Reports
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    • v.45 no.11
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    • pp.671-676
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    • 2012
  • Caloric restriction (CR) has been associated with health benefits and these effects have been attributed, in part, to modulation of oxidative status by CR; however, data are still controversial. Here, we investigate the effects of seventeen weeks of chronic CR on parameters of oxidative damage/modification of proteins and on antioxidant enzyme activities in cardiac and kidney tissues. Our results demonstrate that CR induced an increase in protein carbonylation in the heart without changing the content of sulfhydryl groups or the activities of superoxide dismutase and catalase (CAT). Moreover, CR caused an increase in CAT activity in kidney, without changing other parameters. Protein carbonylation has been associated with oxidative damage and functional impairment; however, we cannot exclude the possibility that, under our conditions, this alteration indicates a different functional meaning in the heart tissue. In addition, we reinforce the idea that CR can increase CAT activity in the kidney. Moreover, CR caused an increase in CAT activity in kidney, without changing other parameters. Protein carbonylation has been associated with oxidative damage and functional impairment; however, we cannot exclude the possibility that, under our conditions, this alteration indicates a different functional meaning in the heart tissue. In addition, we reinforce the idea that CR can increase CAT activity in the kidney.

p53-mediated Inhibitory Mechanism on HIV-1 Tat is Likely to be Associated with Tat-Phosphorylation (p53에 의한 HIV-1 Tat 활성억제와 인산화관련 가능성 연구)

  • Byune, Hee-Sun;Lee, Sang-Gu;Bae, Yong-Soo
    • The Journal of Korean Society of Virology
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    • v.28 no.1
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    • pp.39-52
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    • 1998
  • HIV-1 tat, a strong transactivator, is essential for the HIV-1 replication and AIDS progression. The Tat function is markedly inhibited by human anti-oncogene p53. This work was initiated to identify the p53-associated inhibitory mechanism on tat-mediated transactivation. Inhibitory function of p53 was confirmed by co-transfection of tat-expressing Jurkat cells with LTR-CAT plasmid, or H3T1 cells (LTR-CAT integrated HeLa cells) with different ratio of pSV-tat/pCDNA-p53 plasmids. Results from the direct protein-protein interaction between soluble p53 and tat, and yeast two-hybrid experiments showed that the co-suppression mechanism is unlikely to be due to the direct interaction. CAT activity was not affected by tat in Jurkat cells which were transfected with p53-promoter-CAT or p53-enhancer-CAT, suggesting that the tat-mediated p53 suppression is not directly associated with p53-promoter. Finally, we have tested protein kinase activity in p53-tranfected Jurkat cells, which might phosphorylate HIV-1 tat, resulting in inhibition of tat function. Some of our data lead us to assume that the p53-mediated tat inhibition is likely to be associated with p53-associated, signaling-mediated phosphorylation of tat, resulting in the dysfunction of tat. This study is now under investigation.

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Differential Expression of Arabidopsis Cab Promoters in Organs of Transformed Tobacco Plants (형질전환된 담배식물체의 기관에 따른 Arabidopsis Cab Promoter의 차별적 실험)

  • 홍순조
    • Journal of Plant Biology
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    • v.35 no.3
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    • pp.229-235
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    • 1992
  • Differential expression of the three chlorophyll afb binding (cab) protein gene (cabl, cab2, and cab3) promoters of Arabidopsis thaliana was studied in tobacco plants transformed with cab-CAT (chloramphenicol acetyltransferase) translational fusions. CAT activity was measured to monitor the activities of the cab promoters. The activity of cabi promoter was higher than the other two in transformed tobacco leaves and also in calli and shoots derived from the leaves. Their activities were organ-specific and were the lowest in roots, medium in stems, and the highest in leaves. The relative activity of cabi promoter in stems comparing to it activity in leaves was, however, much higher than the values of cab2 and cab3. When the cab promoter activity was expressed as CAT activity per unit chlorophyll instead of CAT activity per unit protein, the relative cab] promoter activity (stem/leaf) became almost unity. This result suggests that cab2 and cab3 show photosynthetic organ-specificity but cabl does not. Similar result was obtained in the differentiation process of stems and leaves from shoots derived from the transgenic tobacco leaves.leaves.

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Cloning and Sequence Analysis of Two Catechol-degrading Gene Clusters from a Phenol-utilizing Bacterium Pseudomonas putida SM25

  • Jung, Young-Hee;Ka, Jong-Ok;Cheon, Choong-Ⅰll;Lee, Myeong-Sok;Song, Eun-Sook;Daeho Cho;Park, Sang-Ho;Ha, Kwon-Soo;Park, Young-Mok
    • Journal of Microbiology
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    • v.41 no.2
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    • pp.102-108
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    • 2003
  • A 6.1 kb Sph I fragment from the genomic DNA of Pseudomonas putida SM 25 was cloned into the veetor pUC19. The open reading frame of catB was found to consist of 1,122 nucleotides. The sequence alignment of the catB gene products from different kinds of bacteria revealed an overall identity ranging from 40 to 98%. The catC gene contained an open reading frame of 96 codons, from which a protein with a molecular mass of about 10.6 kDa was predicted. The amino acids in the proposed activesite region of CatC were found to be almost conserved, including the charged residues. Since the catBC genes in P. putida SM25 were tightly linked, the could be regulated under coordinate transcription, and transcribed from a single promoter located upstream of the catB gene, as in P. putida RBI.

Solubilities and Activities of Chloramphenicol Acetyltransferase and $\beta$-Lactamase Overproduced by the T7 Expression System in Escherichia coli (대장균에서의 T7 발현체계에 의하여 과잉생산된 클로람페니콜 아세틸전이효소와 베타-락타메이즈의 수용성과 활성)

  • Kim, Han-Bok
    • Korean Journal of Microbiology
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    • v.31 no.4
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    • pp.274-278
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    • 1993
  • Overproduced proteins in many cases result in forming insoluble inclusion bodies, and their formation might be due to high concentration of protein. To investigate how proteins become insoluble, chloramphenicol acetyltransferase (CAT) and .betha.-lactamase were overproduced, and their solubilities and activities were determined. CAT was accumulated from 9 to 45% of total cellular protein in a fully soluble form without inclusion body formation. CAT specific activity was shown to be proportional to the amount of the protein produced. Moderately produced .betha.-lactamase by the phase T7 expression system at 30.deg.C comprised only mature forms in a soluble form. However, overproduced .betha.-lactamase at 37.deg.C became insoluble. Most precursor forms of .betha.-lactamase in the cytoplasm were insoluble, whereas majority of the mature forms in the periplasm space were soluble. Also, chaperone GroE proteins which assist proper protein folding and translocation did not increase .betha.-lactamase solubility significantly under the experimental condition. It seems that the formation of inclusion bodies in the cell is related to the nature of protein itself rather than just to high concentration of protein.

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Development of Genetic System for Isolation of SSU rRNA Mutants that Bypass SecM-Mediated Ribosome Stalling (SecM에서 유래한 접착펩타이드에 의한 라이보솜 정지를 우회하는 SSU rRNA 돌연변이체 발굴을 위한 유전학적 시스템 개발)

  • Ha, Hye-Jeong;Kim, Hong-Man;Yeom, Ji-Hyun;Lee, Kang-Seok
    • Korean Journal of Microbiology
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    • v.44 no.4
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    • pp.271-276
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    • 2008
  • Ribosome stalling by nascent sticky peptide has been reported in several organisms across the kingdom. To test whether small subunit (SSU) rRNA is involved in this phenomenon, we developed a genetic system that utilized the specialized ribosome system to isolate SSU rRNA mutants that enable ribosomes to bypass the SecM-derived sticky peptide in protein synthesis. In this system, CAT-SecM mRNA, which encodes CAT protein containing the sticky peptide derived from SecM, is only translated by specialized ribosomes. These ribosomes were shown to transiently stall on CAT-SecM mRNA followed by the synthesis of the sticky peptide. Expression of specialized ribosomes resulted in the decreased steady-state level of CAT-SecM mRNA, which is consistent with a notion that ribosome stalling induces mRNA degradation. Isolation and characterization of SSU rRNA mutations using this genetic system that are sufficient to circumvent ribosome stalling induced by the SecM-derived sticky peptide will provide evidence of SSU rRNA function in mRNA cleavage.

The Involvement of Protein Kinase C and Tyrosine Kinase in Vanadate-induced Contraction

  • Sim, Sang-Soo;Kim, Chang-Jong
    • Archives of Pharmacal Research
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    • v.21 no.3
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    • pp.315-319
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    • 1998
  • Gastric smooth muscle of cats was used to investigate the involvement of protein kinase in vanadate-induced contraction. Vanadate caused a contraction of cat gastric smooth muscle in a dose-dependent manner. Vanadate-induced contraction was totally inhibited by 2 mM EGTA and 1.5 mM $LACI_3$ and significantly inhibited by $10\mu$M verapamil and $1\mu$M nifedipine, suggesting that vanadate-induced contraction is dependent on the extracellular $Ca^{2+}$ concentration, and the influx of extracellular $Ca^{2+}$ was mediated through voltage-dependent $Ca^{2+}$ channel. Both protein kinase C inhibitor and tyrosine kinase inhibitor significantly inhibited the vanadate-induced contraction and the combined inhibitory effect of two protein kinase inhibitors was greater than that of each one. But calmodulin antagonists did not have any influence on the vanadate-induced contraction. On the other hand, both forskolin ($1\mu$M) and sodium nitroprusside ($1\mu$M) significantly inhibited vanadate-induced contraction. Therefore, these results suggest that both protein kinase C and tyrosino kinase are involved in the vanadate-induced contraction which required the influx of extracellular $Ca^{2+}$ in cat gastric smooth muscle, and that the contractile mechanism of vanadate may be different from that of agonist binding to its specific receptor.

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