한국산 무우 묘목의 성장에 따른 Peroxidase isozyme 들의 조직특이성 발현

이미영,김승수 ( Mi Young Lee,Soung Soo Kim ) 생화학분자생물학회 1990 BMB Reports Vol.23 No.4

Changes in the pattern of isoperoxidases and the specific activity of peroxidases were examined from cotyledone, hypocotyl and root portion of the Korean radish seedlings grown for 3, 5 and 7 days. The specific activity of peroxidases was very high in the root compared with that of hypocotyl and cotyledone portion of the seedlings and peroxidase isozyme patterns in these three organs were changed during entire growth periods. Total cellular RNA was isolated from hypocotyl and root portion of the seedlings with guanidium thiocyanate and guanidine HCl method. The total RNA was further fractionated by oligo (dT)-cellulose chromatography. For translation of peroxidase mRNAs in cell-free protein synthesizing system, rabbit reticulocyte lysate was tested and the optimum translational conditions of peroxidase mRANs were determined. The poly(A) +RNA and poly(A)-RNA isolated from hypocytyl and root portion of the seedlings grown for 5 and 7 days were translated in vitro and the correlation between the changes in the expression pattern of isoperoxidases and qualitative and quantitative changes in peroxidase mRNA populations was investigated. The peroxidase activity of poly(A)-RNAs was much higher than that of poly(A)+RNA in seedlings grown for 7 days when compared with that of poly(A)-RNAs from seedlings grown for 5 days.

한국산 무우로부터 Far Migrating Anionic isoperoxidase A3 의 정제 및 성질

이미영,김승수 ( Mi Young Lee,Soung Soo Kim ) 생화학분자생물학회 1990 BMB Reports Vol.23 No.4

A far migrating anionic isoperoxidase, named A₃, was isolated from Korean radish (Raphanus sativas L.) root. Purification of the enzyme was accomplished by CM-cellulose chromatography, DEAF-Sephacel chromatography and Sephadex G-75 gel filtration. The enzyme was a glycoprotein having molecular weight of approximately 50,000 as determined by SDS-PAGE and Sephadex G-150 gel filtration and the pI value was 3.0. The optimum pH of the enzyme was 6.0 for guaiacol and H₂O₂and the Km values for guaiacol and H₂O₂ were 8 mM and 1 mM, respectively. Substrate studies with natural phenolic compounds were performed and their Km values were determined. In comparison with other Korean radish isoperoxidases, A₃ had higher affinity for esculetin. A particular interest is that the enzyme showed as sigmoidal substrate saturation curve against scopoletin despite of its single Subunit structure.

Tunicamycin 과 2 - deoxy - D - glucose 가 Cellular Peroxidase 와 Secretory Peroxidase 의 생성에 미치는 영향

우구,이미영,김승수 ( Koo Woo,Mi Young Lee,Soung Soo Kim ) 생화학분자생물학회 1987 BMB Reports Vol.20 No.1

The effects of tunicamycin(TM) and 2-deoxy-D-glucose(2-D-G) treatment on the specific activities and isozyme patterns of the cellular and secretory peroxidases of tobacco callus were investigated. The specific activities of the cellular peroxidases were increased by TM and 2-D-G treatment during entire culture periods, whereas the specific activities of the secretory peroxidases were decreased by TM and 2-D-G treatment. Comparisons of the isoperoxidase patterns of TM treated groups and 2-D-G treated groups with those of control groups showed that both TM and 2-D-G cause the significant elevation of the activities of cathodic isoperoxidases, especially isoperoxidase C₄ which is a glycoprotein. But the decrease of the specific activities of the secretory peroxidases by TM and 2-D-G was mainly due to the lowered level of anodic isoperoxidase A₃ which is also a glycoprotein. Immunoprecipitation with anti-A₃antibody and anti-C₄ antibody were carried out to examine the changes in the synthesis and secretion of these isozymes due to the treatment of the callus with the glycosylation inhibitors. These results suggested that the increase of cellular C₄ activity was due to the increased amount of C₄ proteins and the decrease of the activity of secretory isoperoxidase A₃ was caused by the decrease of A₃ proteins in the medium in the presence of the glycosylation inhibitors.

Characteristics of Two Anodic Isoperoxidases from Tobacco Callus

이미영,김정아,김승수,Lee, Mi-Young,Kim, Jeong-A,Kim, Soung-Soo 생화학분자생물학회 1988 한국생화학회지 Vol.21 No.2

담배 조직 배양 (Nicotiana tobacum L. var., Virginia 115)으로부터 두개의 isoperoxidase $A_3$와 $A_4$를 정제하고 이 효소의 몇가지 성질을 조사하였다. 정제된 isoperoxidase $A_3$의 분자량은 SDS-Polyacrylamide gel electrophoresis에서 38,000이었고 Sepadex G-150 gel filtration에 의해서 38,500으로 나타났으며, isoperoxidase $A_4$의 분자량은 SDS-Polyacrylamide gel electrophoresis와 gel filtration에 의해서 모두 43,000이었으므로 이 두 효소는 모두 monomer로 존재한다고 예상된다. Isoperoxidase $A_3$와$A_4$의 guaiacol에 대한 최적 pH는 각각 6.0과 7.0이었고 $K_m$ 값은 이 두 효소 모두 5mM로 동일하였다. Scopoletin, esculetin, ferulic acid와 chlorogenic acid 등의 천연 페놀화합물을 이용한 기질 연구에서 isoperoxidase $A_3$는 각각 0.18 mM, 0.24 mM, 0.26 mM와 0.35 mM의 $K_m$ 값을 나타내었는데 이 값은 isoperoxidase $A_4$의 경우와 크게 다르지 않았다. 또한 이 결과는 다른 담배 조직 배양종인 W-38에서 분리한 $A_3$와 WR-132에서 분리한 $A_f$의 $K_m$ 값과 크게 다르지 않았다. 그러나 이미 scopoletin peroxidase로 그 성질이 특별히 알려져 있는 W-38 isoperoxidase $A_3$와는 달리 Virginia 조직 배양에서 분리한 isoperoxidase $A_3$와 $A_4$는 기질 선호도에 있어서 scopoletin을 특별히 선호하지는 않았다. Two anodic isoperoxidases, named isoperoxidase $A_3$ and $A_4$, were isolated from tobacco callus line, Nicotiana tabaccum L., var. Virginia 115. Isolation of the enzymes was accomplished using DEAE-cellulose ion exchange chromatography and Sephadex G-150 gel filtration. The molecular weight of isoperoxidase $A_3$ was approximately 38,000 as determined by SDS polyacrylamide gel electrophoresis and 38,500 estimated with Sephadex G-150 gel filtration. The molecular weight of isoperoxidase $A_4$ was 43,000 as determined by SDS polyacrylamide gel electrophoresis and Sephadex G-150 gel filtration. The optimum pHs for guaiacol with isoperoxidase $A_3$ and $A_4$ were pH 6.0 and pH 7.0, respectively, and these two isoperoxidases had same $K_m$ value of 5 mM for guaiacol. Substrate studies with natural phenolic compounds such as scopoletin, esculetin, ferulic acid and chlorogenic acid were performed and the $K_m$ values for these substrates were 0.18 mM, 0.24 mM, 0.26 mM and 0.35 mM, respectively, in case of isoperoxidase $A_3$. Such $K_m$ values did not differ greatly from those of isoperoxidase $A_4$ from Virginia strain and known scopoletin peroxidase $A_f$ from WR-132 strain. In comparison with isoperoxidase $A_3$ from W-38 strain and $A_f$ from WR-132 strain which were known as scopoleticn peroxidases, these two enzymes from Virginia strain showed notable differences in scopoletin oxidizing ability in terms of relative substrate oxidation ratios.

담배조직배양으로부터 isoperoxidase 의 분리 및 성질

이미영,김정아,김승수 ( Mi Young Lee,Jeong A Kim,Soung Soo Kim ) 생화학분자생물학회 1988 BMB Reports Vol.21 No.2

Two anodic isoperoxidases, named isoperoxidase A₃ and A₄, were isolated from tobacco callus line, Nicotiana tabaccum L., var. Virginia 115. Isolation of the enzymes was accomplished using DEAE-cellulose ion exchange chromatography and Sephadex G-150 gel filtration. The molecular weight of isoperoxidase A₃ was approximately 38,000 as determined by SDS polyacrylamide gel electrophoresis and 38,500 estimated with Sephadex G-150 gel filtration. The molecular weight of isoperoxidase A₄ was 43,000 as determined by SDS polyacrylamide gel electrophoresis and Sephadex G-150 gel filtration. The optimum pHs for guaiacol with isoperoxidase A₃ and A₄ were pH 6.0 and pH 7.0, respectively, and these two isoperoxidases had same K_m value of 5 mM for guaiacol. Substrate studies with natural phenolic compounds such as scopoletin, esculetin, ferulic acid and chlorogenic acid were performed and the K_m values for these substrates were 0.18 mM, 0.24 mM, 0.26 mM and 0.35 mM, respectively, in case of isoperoxidase A₃. Such K_m values did not differ greatly from those of isoperoxidase A₄ from Virginia strain and known scopoletin peroxidase A_f from WR-132 strain. In comparison with isoperoxidase A₃ from W-38 strain and A_f from WR-132 strain which were known as scopoleticn peroxidases, these two enzymes from Virginia strain showed notable differences in scopoletin oxidizing ability in terms of relative substrate oxidation ratios.

Effect of Tunicamycin and 2-deoxy-D-glucose on the Cellular and Secreted Isoperoxidase Levels of Tobacco Callus

우구,이미영,김승수,Woo, Koo,Lee, Mi-Young,Kim, Soung-Soo Korean Society for Biochemistry and Molecular Biol 1987 한국생화학회지 Vol.20 No.1

담배 callus 배양시 tunicamycin(TM)과 2-deoxy-D-glucose(2-D-G)가 세포내 peroxidase와 배지로 분비되는 peroxidase의 활성과 isozyme pattern에 미치는 영향을 조사하였다. TM과 2-D-G 처리에 의하여 세포내 peroxidase의 비활성도는 전 배양기간동안 현저히 증가한 반면 배지로 분비되는 peroxidase의 비활성도는 배양초기를 제외하고는 점차 감소하였다. Isozyme patterns변화를 조사한 결과 M과 2-D-G처리에 의하여 세포내 peroxidase 중 glycoprotein인 cathodic isoperoxidase $C_4$ 가 뚜렷한 증가를 보인 반면 또 다른 glycoprotein인 anodic isoperoxidase $A_3$는 감소를 나타냈다. 한편 TM과 2-D-G 처리시 배지로 분비되는 peroxidase의 비활성도 감소는 주로 isoperoxidase $A_3$의 감소에 기인한다는 사실도 확인하였다. Anti-$A_3$ 혈청과 anti-$C_4$혈청을 사용하여 isoperoxidase $C_4$와 $A_3$의 양의 변화를 정량한 결과 비활성도 변화 및 pattern 변화와 일치하는 것으로 나타났다. The effects of tunicamycin(TM) and 2-deoxy-D-glucose(2-D-G) treatment on the specific activities and isozyme patterns of the cellular and secretory peroxidases of tobacco callus were investigated. The specific activities of the cellular peroxidases were increased by TM and 2-D-G treatment during entire culture periods, whereas the specific activities of the secretory peroxidases were decreased by TM and 2-D-G treatment. Comparisons of the isoperoxidase patterns of TM treated groups and 2-D-G treated groups with those of control groups showed that both TM and 2-D-G cause the significant elevation of the activities of cathodic isoperoxidases, especially isoperoxidase $C_4$ which is a glycoprotein. But the decrease of the specific activities of the secretory peroxidases by TM and 2-D-G was mainly due to the lowered level of anodic isoperoxidase $A_3$ which is also a glycoprotein. Immunoprecipitation with anti-$A_3$ antibody and anti-$C_4$ antibody were carried out to examine the changes in the synthesis and secretion of these isozymes due to the treatment of the callus with the glycosylation inhibitors. These results suggested that the increase of cellular $C_4$ activity was due to the increased amount of $C_4$ proteins and the decrease of the activity of secretory isoperoxidase $A_3$ was caused by the decrease of $A_3$ proteins in the medium in the presence of the glycosylation inhibitors.

천식환자에서 병발된 S 단백 결핍과 연관된 폐동맥 색전증

박정화 ( Chung Hwa Park ),윤승배 ( Seung Bae Yoon ),김대범 ( Dae Bum Kim ),이한희 ( Han Hee Lee ),김찬준 ( Chan Joon Kim ),김정호 ( Jeong Ho Kim ),고훈영 ( Hoon Young Ko ),이미정 ( Mi Jeong Lee ),이화정 ( Hwa Jeong Lee ),김승수 ( 대한천식알레르기학회 2010 천식 및 알레르기 Vol.30 No.2

Asthma is a chronic multifactorial disease with frequent exacerbations in many patients. Much effort has been made to determine prevalent factors associated with difficult- to-control asthma. Here we report the case of a 57-year-old man who was treated for asthma during the last 4 years at our pulmonology clinic. Recently, the patient experienced severe dyspnea for more than 3 months despite the addition of the leukotriene modifier, theophylline and oral cortic- osteroid. Chest CT scans revealed multifocal areas of pulmonary arterial thrombosis. Doppler ultrasonograms showed multiple deep vein thrombi. The patient was diagnosed of having protein S deficiency and was treated with low molecular weight heparin, followed by oral warfarin. Now, he has no pulmonary arterial thrombosis and has no more asthmatic attack for half a year. (Korean J Asthma Allergy Clin Immunol 2010;30:135-139)

Tunicamycin 과 2 - deoxy - D - glucose 가 Cellular Peroxidase 와 Secretory Peroxidase 의 생성에 미치는 영향

이미영,김승수,우구 생화학분자생물학회 1992 BMB Reports Vol.14 No.2

The effects of tunicamycin(TM) and 2-deoxy-D-glucose(2-D-G) treatment on the specific activities and isozyme patterns of the cellular and secretory peroxidases of tobacco callus were investigated. The specific activities of the cellular peroxidases were increased by TM and 2-D-G treatment during entire culture periods, whereas the specific activities of the secretory peroxidases were decreased by TM and 2-D-G treatment. Comparisons of the isoperoxidase patterns of TM treated groups and 2-D-G treated groups with those of control groups showed that both TM and 2-D-G cause the significant elevation of the activities of cathodic isoperoxidases, especially isoperoxidase C₄ which is a glycoprotein. But the decrease of the specific activities of the secretory peroxidases by TM and 2-D-G was mainly due to the lowered level of anodic isoperoxidase A₃ which is also a glycoprotein. Immunoprecipitation with anti-A₃antibody and anti-C₄ antibody were carried out to examine the changes in the synthesis and secretion of these isozymes due to the treatment of the callus with the glycosylation inhibitors. These results suggested that the increase of cellular C₄ activity was due to the increased amount of C₄ proteins and the decrease of the activity of secretory isoperoxidase A₃ was caused by the decrease of A₃ proteins in the medium in the presence of the glycosylation inhibitors.