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쇠 뇌에서 정제된 Alkaline Ribonuclease 와 Ribonuclease Inhibitor 의 특성
김금이,조성우 ( Geum Yi Kim,Sung Woo Cho ) 생화학분자생물학회 1991 BMB Reports Vol.24 No.3
An alkaline ribonuclease (pH optimum 8) has been purified in a five step procedure and a ribonuclease inhibitor (pH optimum 7.6) has been purified by affinity chromatography on Sepharose-RNase A from bovine brain. The purified proteins appear homogeneous on SDS-polyacrylamide gel electrophoresis. The data from gel filtration and SDS-polyacrylamide gel electrophoresis indicate a molecular weight of 27,000 for alkaline ribonuclease and 51,000 for the inhibitor. By gel filtration, the molecular size of the ribonuclease-inhibitor complex is 75,000, indicative of 1:1 binding on a molecular basis. The inhibition of the action of alkaline ribonuclease on yeast RNA by the inhibitor is noncompetitive with a K_i of 0.9 nM. The alkaline ribonuclease activity either in crude extracts or through the purification steps was affected by p-chloromercuribenzoic acid. Essentially no free alkaline ribonuclease activity could be detected in bovine brain homogenate in the absence of p-chloromercuribenzoic acid. Inactivation of the inhibitor by sulfhydryl reagents causes the dissociation of the ribonuclease-inhibitor complex into active ribonuclease and inactive inhibitor. Blocking of lysine residue(s) of the alkaline RNase by a chemical modification inactivated the RNase activity, and interrupted a formation of RNase-inhibitor complex. It indicates that the lysine residue(s) of the alkaline RNase might be crucial not only for the alkaline RNase activity but also for the binding of the inhibitor to the enzyme.
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사람 혈소판에서 분리한 Transforming Growth Factor - β가 초회 배양된 Mesangial 세포의 복제 및 Collagen 합성에 미치는 영향
조성우,박수길,김금이,이희봉,이재담 ( Sung Woo Cho,Su Kil Park,Geum Yi Kim,Hee Bong Lee,Jae Dam Lee ) 생화학분자생물학회 1992 BMB Reports Vol.25 No.3
The effect of TGF-β on cell replication and collagen synthesis in primarily cultured mesangial cells from rats was studied in vitro. The TGF-β was purified from human platelets by acidic ethanol extraction and gel filtration with FPLC. The effect of TGF-β on the DNA synthesis and the collagen synthesis in mesangial cells depended on both TGF-β concentration and cell density in monolayer culture. Concentrations of TGF-β ranging from 0.1∼15 ng/ml were evaluated for their effects upon DNA synthesis. In confluent cells, when compared to the growth rate in standard medium, TGF-β concentrations in the range 0.1∼15 ng/ml reduced a cell proliferation up to 35%, suggesting inhibition of cell division. The effect of TGF-β on collagen synthesis in mesangial cells was biphasic. In confluent cells, the addition of TGF-β induced a 2-fold increase in the production of collagen when compared with cells grown in standard medium. The maximal effect of TGF-β occurred at 0.5 ng/ml. At higher concentrations there was a decrease in the action of TGF-β with the effects on collagen production. In subconfluent cells, however, TGF-β showed no significant effects on the DNA synthesis and the collagen synthesis at all concentrations. These results support that TGF-β may play an important role as a multifunctional regulator in the pathogenesis of glomerulosclerosis. v
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조성우,박수길,김금이,이희봉,이재담,Cho, Sung-Woo,Park, Su-Kil,Kim, Geum-Yi,Lee, Hee-Bong,Lee, Jae-Dam 생화학분자생물학회 1992 한국생화학회지 Vol.25 No.3
쥐의 mesangial 세포를 초회 배양하여 Transforming Growth Factor-$\beta$(TGF-$\beta$)가 세포의 복제 및 collagen 합성에 미치는 영향을 조사하였다. TGF-$\beta$는 사람 혈소판에서 산성 에탄올 침전법 및 FPLC를 이용한 젤 크로마토그래피의 방법으로 정제되었다. TGF-$\beta$가 mesangial 세포를 복제 및 collagen 합성에 미치는 영향은 TGF-$\beta$ 농도와 세포의 밀도에 따라 달라졌다. Confluent 세포의 경우 0.1~15 ng/ml의 농도에서 TGF-$\beta$는 mesangial 세포의 복제를 35%까지 감소시켰다. Collagen 합성은 0.5 ng/ml의 TGF-$\beta$ 농도에서는 2배로 증가되었으나 그 이상의 농도에서는 감소하기 시작하는 두 가지의 조절 양상을 보였다. 그러나 subconfluent 세포의 경우에는 세포의 복제 빛 collagen 합성 모두에 TGF-$\beta$는 아무런 효과를 나타내지 못하였다. 본 연구결과는 TGF-$\beta$가 glomerulosclerosis에 있어서 중요한 역할을 하리라는 가능성을 제시해주고 있다. The effect of TGF-$\beta$ on cell replication and collagen synthesis in primarily cultured mesangial cells from rats was studied in vitro. The TGF-$\beta$ was purified from human platelets by acidic ethanol extraction and gel filtration with FPLC. The effect of TGF-$\beta$ on the DNA synthesis and the collagen synthesis in mesangial cells depended on both TGF-$\beta$ concentration and cell density in monolayer culture. Concentrations of TGF-$\beta$ ranging from 0.1~15 ng/ml were evaluated for their effects upon DNA synthesis. In confluent cells, when compared to the growth rate in standard medium, TGF-$\beta$ concentrations in the range 0.1~15 ng/ml reduced a cell proliferation up to 35%, suggesting inhibition of cell division. The effect of TGF-$\beta$ on collagen synthesis in mesangial cells was biphasic. In confluent cells, the addition of TGF-$\beta$ induced a 2-fold increase in the production of collagen when compared with cells grown in standard medium. The maximal effect of TGF-$\beta$ occurred at 0.5 ng/ml. At higher concentrations there was a decrease in the action of TGF-$\beta$ with the effects on collagen production. In subconfluent cells, however, TGF-$\beta$ showed no significant effects on the DNA synthesis and the collagen synthesis at all concentrations. These results support that TGF-$\beta$ may play an important role as a multifunctional regulator in the pathogenesis of glomerulosclerosis.
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기술보고 : 관입형 피조미터와 시피지미터를 이용한 안성천 상류구간 하상 수리전도도 측정
이정우 ( Jeong Woo Lee ),전선금 ( Seon Geum Chun ),이명재 ( Myeong Jae Yi ),김남원 ( Nam Won Kim ),정일문 ( Il Moon Chung ),이민호 ( Min Ho Lee ) 대한지질공학회 2015 지질공학 Vol.25 No.3
안성천에 위치한 공도 수위표 상류 구간에 대해 하상퇴적층을 통과하는 하천수-지하수 상호교환량과 수리경사를 측정하여 하상퇴적층의 수리전도도를 산정하였다. 하천수-지하수 상호유동량은 자체 제작한 시피지 미터를 이용하여 측정하였고, 수리경사는 장심도와 단심도 두 가지 종류의 관입형 피조미터를 하상퇴적층에 설치하여 피조미터내 수두 차이와 관입깊이 차이를 측정하여 산정하였다. 하천수-지하수 교환량은 -1.55 × 10-6~1.77 × 10-5 m/s, 연직방향 수리경사는 -0.122에서 0.030로 측정되었다. 교환량과 수리경사로 부터 연직방향 수리전도도를 산정한 결과 1.77 × 10-5~1.97 × 10-3 m/s로 지점별로 큰 차이를 나타내었다. 측정 결과는 Calver (2001)가 제시한 하상퇴적물 수리전도도의 일반적인 범위내에서 분포하였으며, 모래, 자갈층이 잘 발달되어 있어 기존의 국내 측정 결과 사례에 비해 큰 값을 보이는 것으로 분석되었다. 본 논문에서 적용한 고강도 스테인레스 재질의 피조미터와 시피지미터를 이용하여 하상퇴적층의 수리전도도를 측정하는 방법은 내구성과 휴대성이 우수하여 실무에서 매우 유용하게 활용될 수 있을 것으로 기대된다. Streambed hydraulic conductivity along the upper reaches of the Gongdo stage of Anseong Stream was estimated through measurements of stream-aquifer exchange rates (using a seepage meter) and vertical hydraulic gradients (using a manually driven piezometer). From the measured data, it was found out that the stream-aquifer exchange rates varied from -1.55 × 10-6 to 1.77 × 10-5 m/s, the corresponding vertical hydraulic gradient varied from -0.122 to 0.030, and the values of the streambed vertical hydraulic conductivity were estimated from 1.77 × 10-5 to 1.97 × 10-3 m/s, with variations representing local differences. The results are within the general range of streambed hydraulic conductivity values suggested by Calver (2001) and are slightly higher than values previously measured at other stream sites in Korea. The combined use of a drive-point piezometer and seepage meter (both constructed of high-strength stainless steel) is expected to be of practical use in the estimation of streambed hydraulic conductance, given the durability and portability of the instruments.
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