• 제목/요약/키워드: Cell-free protein synthesis

검색결과 78건 처리시간 0.022초

A PROMISING NEW ANTI-WRINKLE INGREDIENT: Pericarpium castaneae extracts

  • Kim, Beom-Jun;Jo, Byoung-Kee;Kim, Jeong-Ha
    • 대한화장품학회지
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    • 제25권4호
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    • pp.57-63
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    • 1999
  • Pericarpium castaneae extracts have variously potent activities, such as anti-oxidative activity and free radical scavenging activity. in vivo and in vitro studies both indicate that pericarpium castaneae extracts acts as a free radical scavenger($IC_{50}:7.6{\mu}g/ml$) stronger than gallic acid($IC_{50}:12.5{\mu}g/ml$) and ellagic acid($IC_{50}:15{\mu}g/ml$) which could prevent cutaneous UV damages and skin aging. The extracts showed a good effect as a anti-oxidant($IC_{50}:50{\mu}g/ml$). It was shown that the appearance of wrinkle in human skin was reduced by topical application of pericarpium castaneae extracts. And the treatment of human skin with the extracts increased the elasticity and moisture of the skin. We investigated the effect of the pericarpium castaneae extracts on production of extracellular matrix using cultured A431 fibroblast cells. The results indicated that the extracts had no detectable effect on collagen synthesis. But synthesis of cell adhesion protein was increased by the extracts. The results suggest that increase of cell adhesion protein synthesis by pericarpium castaneae extracts has closely related to reduction of wrinkle in skin.

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A PROMISING NEW ANTI-WRINKLE INGREDIENT : Pericarpium castaneae extracts

  • Kim, Beom-Jun;Jo, Byoung-Kee;Kim, Jeong-Ha
    • 대한화장품학회:학술대회논문집
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    • 대한화장품학회 1999년도 IFSCC . ASCS 학술대회 발표 논문
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    • pp.57-64
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    • 1999
  • Pericarpium castaneae extracts have variously potent activities, such as anti-oxidative activity and free radical scavenging activity. in vivo and in vivo studies both indicate that pericarpium castaneae extracts acts as a flee radical scavenger ($IC_{50}$/: 7.6$\mu\textrm{g}$/ml) stronger than gallic acid($IC_{50}$/: 12.5$\mu\textrm{g}$/ml) and ellagic acid($IC_{50}$/: 15$\mu\textrm{g}$/ml) which could prevent cutaneous UV damages and skin aging. The extracts showed a good effect as a anti-oxidant ($IC_{50}$/: 50$\mu\textrm{g}$/ml). It was shown that the appearance of wrinkle in human skin was reduced by topical application of pericarpium castaneae extracts. And the treatment of human skin with the extracts increased the elasticity and moisture of the skin. We investigated the effect of tile pericarpium castaneae extracts on production of extracellular matrix using cultured A431 fibroblast cells. The results indicated that the extracts had no detectable effect on collagen synthesis, But synthesis of cell adhesion protein was increased by the extracts. The results suggest that increase of cell adhesion protein synthesis by pericarpium castaneae extracts has closely related to reduction of wrinkle in skin.

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INTRACELLULAR AMINO ACID PROFILE OF RUMEN BACTERIA AS INFLUENCED BY UREA FEEDING AND ITS DURATION

  • Kobayashi, Y.;Wakita, M.;Hoshino, S.
    • Asian-Australasian Journal of Animal Sciences
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    • 제6권4호
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    • pp.619-622
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    • 1993
  • Rumen bacterial amino acids in sheep on urea diet were monitored to assess a possible change in amino acid synthesis as a long term response to high rumen ammonia environment. A sheep was fed a semipurified diet with soybean meal, followed by a diet with urea as a main nitrogen source. Mixed rumen bacteria were harvested from ruminal fluid taken 3 h after feeding (twice in soybean meal feeding and 6 times in urea feeding) and fractionated as cell wall, proteins and protein-free cell supernatant of monitor amino acids in each fraction. Ruminal ammonia concentration at the sampling ranged from 5.7 to 39.5 mgN/dl. Cell wall and protein fractions of mixed rumen bacteria were stable in their amino acid composition regardless of nitrogen sources of diet and the feeding duration. However, protein-free cell supernatant fraction showed a higher alanine proportion with urea feeding (18.6 and 28.2 molar % of alanine for samples from sheep fed soybean meal and urea, respectively) and its duration (20.6 and 32.9 molar % for samples from sheep on urea diet for 1 and 65 days, respectively). Total free amino acid level of bacteria was depressed in the initial period of urea feeding but restored on 65th day of the feeding. These results suggest that an alanine synthesizing system may develop in rumen bacteria as urea feeding becomes longer.

Chlorella 의 물질대사에 미치는 미양원소의 결핍효과(제 2 ) -, 리보 및 의 생합성능에 관하여- (Effect of micronutritional-element deficiencies on the metabolism of Chlorella cells. (II) On the biosynthetic activities of protein, nucleic acids and phospholipid)

  • 이영록;진평;심웅섭
    • 미생물학회지
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    • 제6권1호
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    • pp.22-28
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    • 1968
  • Chlorella ellipsoidea cells were cultured in an iron, copper, zinc, manganese, molybdenum or boron-free medium. Biosynthetic activities of nucleic acids, protein and phospholipid in chlorella cells, which were growing in a microelement deficient medium were compared with those of the normal cells by measuring the contents of phosphate, amino acids or UV-absorbing substances in the various cell fractions. When the algae were grown in a molybdenum-free medium, the amounts of phosphate in the acid-soluble fraction of the cells increased, whereas the amounts of alkali-stable protein and RNA decreased compared with the normal cells showing that the synthesis of protein and RNA from the early products of photosynthesis was inhibited. When the algae were grown in a boron-free medium, amounts of alkali-labile protein and phospholipid of the cells decreased, while the amount of phosphate in acid-soluble fraction increased compared with the normal cells showing that the biosynthesis of protein and phospholipid from the early products of photosynthesis was retarded. In general, amounts of protein and RNA in the microelement deficient cells significantly decreased compared with those of the normal cells. Phosphate content in the acid-soluble fraction of the algal cell grown in an zinc, copper, molybdenum, or boron-free medium increased considerably, whereas that of the algal cell grown in an iron or manganese-free medium decreased remarkably compared with that of the control. It is considered, therefore, that molybdenum, zinc, copper and boron etc. play an important role in the biosyntbesis of macromolecule from acid-soluble phosphate compounds, in contrast to the principal action of iron and manganese on the photosynthetic process itself.

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해당과정의 활성화를 통한 무세포 단백질 발현 시스템에서의 ATP 재생 (Regeneration of ATP through an Activated Glycolytic Pathway in a Cell-free Extract and its Application for Protein Expression)

  • 김동명;금정원;김태완;오인석;최차용
    • KSBB Journal
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    • 제19권6호
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    • pp.467-470
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    • 2004
  • 해당 작용의 중간체를 에너지원으로 이용한 무세포 단백질 발현 반응에서의 낮은 재현성 및 단백질 생산성은 반응액의 pH 및 NAD의 존재에 의해 크게 영향을 받는다는 사실을 밝혀내었다. 기존의 PEP를 사용하는 표준반응 용액에서 PEP를 G-6-P로 대체하고 동시에 반응액의 pH 및 NAD 농도를 최적화 함으로써 반응액 1 mL당 약 $300{\mu}g$에 이르는 단백질을 회분식 반응으로 발현할 수 있었다. ATP 재생 방법의 개선을 통한 회분식 무세포 단백질 발현의 생산성 향상은 다종 유전자의 고속 번역을 통한 기능 규명에 있어서 유용한 도구로서 사용될 수 있을 것으로 기대된다.

An In Vitro Assay to Screen for Translation Inhibitors

  • Song, Chin-Hee;Paik, Hyoung-Rok;Seong, Chi-Nam;Choi, Sang-Ki
    • Journal of Microbiology and Biotechnology
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    • 제16권10호
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    • pp.1646-1649
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    • 2006
  • Protein synthesis is the ultimate outcome of gene expression which, in turn, is regulated by several translation factors. We attempted to identify substances that can inhibit the translation process in vitro when the outcome protein is luciferase. To this end, we developed a sensitive cell-free protein synthesis assay using luciferase as the reporter. The synthesis of luciferase increased proportionately as mRNA was added to a $15-{\mu}l$reaction medium in concentrations raging from 5 ng to 500 ng. The maximum amount of luciferase was synthesized when the media were incubated at $25^{\circ}C$ for 40 min. The concentration of each compound that inhibited luciferase production by 50% ($IC_{50}$) was calculated. Hygromycin, puromycin, and cycloheximide yielded an $IC_{50}$ of 0.008, 0.8, and $0.7{\mu}g/ml$, respectively. A filtrate of Streptomyces spp. isolates inhibited protein synthesis up to S-fold when added to the in vitro translation assay mixture.

Chronic Treatment of Ethanol Inhibits Proliferation of Normal Fibroblasts, but Not Oncogenic ras-Transformed Cells

  • Gu, Young-Hwa;Park, Mi-Sun;Jhun, Byung-H.
    • Biomolecules & Therapeutics
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    • 제6권4호
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    • pp.345-350
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    • 1998
  • The adverse effects of ethanol on cell proliferation have been described for a variety of tissues and cells. In the present study, we investigated whether chronic ethanol intoxication impairs the cell proliferation and DNA synthesis induced by oncogenic $H-ras^{V12}$ - and $v-K-ras^{V12}$-transformed cells. Ethanol treatment inhibited the cell proliferation and the DNA synthesis of control parental fibroblasts in a time- and dose-dependent manner. In contrast, ethanol did not suppress the proliferation of either oncogenic $H-ras^{V12}$ - or $v-K-ras^{V12}$ -transformed fibroblasts. Microinjection of oncogenic $H-Ras^{V12}$ protein induces DNA synthesis and ethanol treatment did not interfere with the DNA synthesis. The antiproliferative toxicity of ethanol was rescued by antioxidants, such as N-acetylcysteine and 4-methlpyrazole. These results indicate that the antiproliferative action site of ethanol toxicity lies upstream or is independent of Ras and ethanol exerts its toxicity through a free radical formation.

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Regulation of Protein Degradation by Proteasomes in Cancer

  • Jang, Ho Hee
    • Journal of Cancer Prevention
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    • 제23권4호
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    • pp.153-161
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    • 2018
  • Imbalance of protein homeostasis (proteostasis) is known to cause cellular malfunction, cell death, and diseases. Elaborate regulation of protein synthesis and degradation is one of the important processes in maintaining normal cellular functions. Protein degradation pathways in eukaryotes are largely divided into proteasome-mediated degradation and lysosome-mediated degradation. Proteasome is a multisubunit complex that selectively degrades 80% to 90% of cellular proteins. Proteasome-mediated degradation can be divided into 26S proteasome (20S proteasome + 19S regulatory particle) and free 20S proteasome degradation. In 1980, it was discovered that during ubiquitination process, wherein ubiquitin binds to a substrate protein in an ATP-dependent manner, ubiquitin acts as a degrading signal to degrade the substrate protein via proteasome. Conversely, 20S proteasome degrades the substrate protein without using ATP or ubiquitin because it recognizes the oxidized and structurally modified hydrophobic patch of the substrate protein. To date, most studies have focused on protein degradation via 26S proteasome. This review describes the 26S/20S proteasomal pathway of protein degradation and discusses the potential of proteasome as therapeutic targets for cancer treatment as well as against diseases caused by abnormalities in the proteolytic system.