Hyperthermia에 있어서 암세포막의 Ion 수송에 관한 연구

장정순,Chang, Chung-Soon 생화학분자생물학회 1982 한국생화학회지 Vol.15 No.4

Tumor cell undergo a rapid change in their K ion content as they are incubated several degrees above the physiological temperature. The K content of P-815 mouse mastocytoma cells decreases while efflux of the ions increases. The ouabain sensitive pumping rate for $^{86}Rb$ suffers a reduction of 40% within an hour. On the other hand, the mouse red blood cells do not indicate any change in the rate of $^{86}Rb$ influx over a 60 min incubation at $43^{\circ}C$. The change in the Na content is not detectable either by the flame emission or by isotope counting. The Na influx, however, increases somewhat soon after the temperature is shifted to $43^{\circ}C$. Since the K content decreases much more than the Na content increases, the net cation content in the cells decrease. This ionic imbalance should result in negatively charged cells unless the culls also lose anions or accumulate other canons.

화분의 Branched-Chain Amino Acid Aminotransferase에 관한 연구(I) 소나무 화분(Pinus densiflora Siebold)의 Aminotransferase에 대하여

장정순,하창우,Chang, Chung-Soon,Ha, Chang-Woo 생화학분자생물학회 1977 한국생화학회지 Vol.10 No.4

소나무 화분으로 부터 branched-chain 아미노산 아미노기 전위 효소 [EC 2. 6. 1. 6]를 검출하기 위하여 화분의 마쇄액을 분별 원심분획한 결과 전체 효소 활성도의 86%가 세포질 분획에서 측정되었고 상기 효소를 정제하기 위하여 DEAE-cellulose column chromatogaphy를 행한 결과 유출용 인산엽 완충액의 농도 180 및 200mM에서 효소 II와 III이 각각 검출되었다. 상기 소나무 화분의 효소 II와 III은 정상 백서의 간 및 뇌 조직에서 추출 정제된 효소와 같은 농도의 유출용 완충액에서 검출된 점으로 미루어 상기 효소들의 성상이 같은 것으로 생각되며 소나무 화분의 경우 효소 II의 비활성도는 III에 비하여 약 2.5배의 활성도를 보였고 이 효소 II 및 III은 chromatography 결과 각각 150배 및 125배로 정제되었다 A study was undertaken on the distribution of branched-chain amino acid aminotransferase (EC 2. 6. 1. 6.) in pine pollen. About 86% of the total enzyme activity was recovered in cytosol fraction, and further purification was carried out with DEAE-cellulose column chromatography. It was demonstrated that there were two types of isozymes in pollen cytosol fraction such as Enzyme II and Enzyme III. One enzyme (enzyme II) was eluted by 180mM phosphate buffer from a DEAE-cellulose column and was specific for leucine. The other enzyme (enzyme III) was eluted by 200mM phosphate buffer and catalyzed the transamination of all three amino acids. The chromatographic elution profile of the enzymes II and III from the pine pollen was very similar with those of normal rat liver and brain. It was found that the enzyme activity was higher in enzyme II than enzyme III. The enzyme II and III were purified about 150 and 125-folds increases in specific activities after chromatography on DEAE-cellulose. The properties of these enzymes are currently under investigation.

Isolation and Chemical Properties of the Surface-Orientated Major Glycoprotein from Bovine Erythrocyte Membrane

장정순,Chang, Chung-Soon 생화학분자생물학회 1977 한국생화학회지 Vol.10 No.4

The major glycoprotein of bovine erythrocyte membrane was isolated after the membrane was solubilized with synthetic fatty acyl ester of palmitoyl-DL-carnitine. The major glycoprotein was purified by calcium phosphate gel treatment, hydroxyapatite, and finally DEAE-cellulose column chromatography. The purified major glycoprotein which was completely water-soluble was a glycolipoprotein. It appeared to be homogeneous and monomeric form. There was not any detectable subunit formation when subjected to polyacrylamide gel electrophoresis in the presence of reducing agents or detergents. This major glycoprotein contains high proportion of sialic acid (43%) and acidic amino acids. The results of crossed-immunoelectrophoresis with rabbit antibody to bovine erythrocyte membrane or intact bovine RBC-depleted antibody clearly indicated that the major glycoprotein was located at the outer surface of the membrane. 소 적혈구 막을 DL-palmitoyl carnitine으로 완전 용해한 뒤 calcium phosphate gel 처리후 hydroxyapatite 및 DEAE-cellulose column chromatography로 major sialoglycoprotein을 분리 정제하였으며 정제된 major sialoglycoprotein은 SDS-polyacrylamide gel electrophoresis 에서 순수한 monomer의 형태로 분리되었다. 정제된 major sialoglycoprotein은 glycolipoprotein으로 소량의 인지질과 cholesterol을 함유하고 있었고, sialic acid의 함량이 높았으며 (43%) 아미노산 조성으로 미루어 산성 단백질이었다. 다음 소적혈구 막에 대한 항혈청 및 소적혈구로 흡수된 각각의 항혈청과 정제된 major sialoglycoprotein을 항원으로 하여 crossed-immunoelectrophoresis를 한 결과 상기 glycoprotein은 당 잔기의 일부가 적혈구막 밖으로 노출되어 있었음을 알 수 있었다.

화분의 Branched - Chani Amino Acid Aminotransferase 에 관한 연구 ( 1 ) 소나무 화분 ( Pinus Densiflora Siebold ) 의 Aminotransferase 에 대하여

장정순,하창우 ( Chung Soon Chang,Chang Woo Ha ) 생화학분자생물학회 1977 BMB Reports Vol.10 No.4

A study was undertaken on the distribution of branched-chain amino acid aminotransferase (EC 2. 6. 1. 6. ) in pine pollen. About 86~ of the total enzyme activity was recovered in cytosol fraction, and further purification was carried out with DEAE-cellulose column chromatography. It was demonstrated that there were two types of isozymes in pollen cytosol fraction such as Enzyme II and Enzyme III. One enzyme (enzyme II) was eluted by 180mM phosphate buffer from a DEAE-cellulose column and was specific for leucine. The other enzyme(enzyme III) was eluted by 200mM phosphate buffer and catalyzed the transamination of all three amino acids. The chromatographic elution profile of the enzymes II and III from the pine pollen was very similar with those of normal rat liver and brain. It was found that the enzyme activity was higher in enzyme II than enzyme III. The enzyme II and III were purified about 150 and 125-folds increases in specific activities after chromatography on DEAE-cellulose. The properties of these enzymes are currently under investigation.

Hyperthermia 에 있어서 암세포막의 Ion 수송에 관한 연구

장정순 ( Chung Soon Chang ) 생화학분자생물학회 1982 BMB Reports Vol.15 No.4

Tumor cell undergo a rapid change in their K ion content as they are incubated several degrees above the physiological temperature. The K content of P-815 mouse mastocytoma cells decreases while efflux of the ions increases. The ouabain sensitive pumping rate for ^(86)Rb suffers a reduction of 40% within an hour. On the other hand, the mouse red blood cells do not indicate any change in the rate of ^(86)Rb influx over a 60 min incubation at 43℃. The change in the Na content is not detectable either by the flame emission or by isotope counting. The Na influx, however, increases somewhat soon after the temperature is shifted to 43℃. Since the K content decreases much more than the Na content increases, the net cation content in the cells decrease. This ionic imbalance should result in negatively charged cells unless the culls also lose anions or accumulate other canons.

소 적혈구 막 Major Glycoprotein 의 분리 정제 및 성상에 관한 연구

장정순 ( Chung Soon Chang ) 생화학분자생물학회 1977 BMB Reports Vol.10 No.4

The major glycoprotein of bovine erythrocyte membrane was isolated after the membrane was solubilized with synthetic fatty acyl ester of palmitoyl-DLcarnitine. The major glycoprotein was purified by calcium phosphate gel treatment, hydroxyapatite, and finally DEAE-celluiose column chromatography. The purified major glycoprotein which was completely water-soluble was a glycolipoprotein. It appeared to be homogeneous and monomeric form. There was not any detectable subunit formation when subjected to polyacrylamide gel electrophoresis in the presence of reducing agents or detergents. This major glycoprotein contains high proportion of sialic acid (43 jb) and acidic amino acids. The results of crossed-immunoelectrophoresis with rabbit antibody to bovine erythrocyte membrane or intact bovine RBC-depleted antibody clearly indicated that the major glycoprotein was located at the outer surface of the membrane.

Swamp eel (Monopterus albus)혈장 Albumin분획의 정제 및 물리화학적 성상에 관한 연구

김상복,장정순,Kim, Sang-Bog,Chang, Chung-Soon 생화학분자생물학회 1982 한국생화학회지 Vol.15 No.4

드렁허리(Monopterus albus)로부터 albumin분획 만을 정제하기 위하여 ammonium sulfate로 crude albumin분획을 얻은 후, 다시 DEAE-Sephadex A-50을 이용한 column chromatography로 albumin분획을 정제하였다. 정제된 albumin분획의 물리화학적 성상을 알기 위하여 organic cation인 rivanol을 이용한 albumin분획의 확인실험, $NaDodSO_4$-PAGE에 의한 분자량 측정, 흡광계수, 침강계수 및 -SH 함량등을 측정하였다. 즉 어류의 혈장으로부터 albumin분획을 분리할때 완만한 구배농도의 증가와 많은 양의 buffer로 유출시켰을 때 정제된 albumin을 얻을 수 있었고 드렁허리 정제 albumin의 분자량은 이미 알려진 다른 동물의 것에 비하여 분자량이 절반 정도였으며 -SH함량도 0.04로 매우낮았다. 끝으로, 흡광계수는 30,320 $mol^{-1}cm^{-1}$이었으며 침강계수는 dimer의 상태일때 4.5 S, neutral sugar의 함량은 대략$39{\pm}0.2{\mu}g$per mg 단백질을 나타내었다. Albumin fraction was purified from the swamp eel by salting out with ammonium sulfate and following by DEAE-Sephadex A50 column chromatography. The purified albumin fraction appeared to be homogeneous by Davis and $NaDodSO_4$ polyacrylamide gel electrophoresis after albumin fraction was treated with organic cation, rivanol (2-ethoxy-6,9, diaminoacridine lactate). The physico-chemical properties on purified albumin fraction were characterized by means of molecular weight determination, -SH content, extinction and sedimentation coefficient, etc.

황복 , Takifugu obscurus , 초기 단계의 소화효소 변화

손규희(Kyu Hee Son),한경남(Kyung Nam Han),장정순(Chung Soon Chang) 한국수산과학회 2001 한국수산과학회지 Vol.34 No.6

황복의 소화효소의 변화과정을 부화직후부터 부화 후 65일까지 측정하였다. 발육단계에 따른 소화효소의 발현과 활성의 변화에서 α-amylase 비활성은 전장 10 ㎜에서 0.0493 U/㎎의 최소값을 나타낸 후, 전장 19 ㎜를 전후하여 0.1480 U/㎎의 최대값을 나타냈다. Trypsin과 pepsin 비활성은 전장 16 ㎜에서 각각 0.0264 U/㎎, 0.0258 U/㎎ 와 전장 24 ㎜에서 0.0178 U/㎎, 0.0201 U/㎎의 값을 가지는 두 번의 peak를 보였고, 이 시기에 황복 자치어의 성장률도 증가하는 경향을 보였다. 또한, trypsin과 pepsin의 비활성을 비교하여 보면, 자어기인 전장 4∼5 ㎜와 치어 II기인 전장 19∼24 ㎜에서는 pepsin이 높았고, 치어 I 기인 전장 11∼16 ㎜와 유어기인 전장 27 ㎜ 이후에서는 trypsin이 높았다. The digestive enzyme activities such as α-amylase, trypsin and pepsin from the laboratory-reared river puffer, Takifugu obscurus, were measured from the time course of 1 day until the 65 day after hatching. In the case of α-amylase, it was showed minimum activity of 0.0493 U/㎎ at the total length (TL) 10 ㎜, and showed maximum activity of 0.1480 U/㎎ at 19 ㎜TL. Trypsin and pepsin were showed their maximum activities of 0.0264, 0.0258 U/㎎ and 0.0178, 0.0201 U/㎎ when the total length of 16 and 24 ㎜, and represented remarkable correlations between the changes of enzyme activity and growth rate. The ontogenetic variations of digestive enzymes were represented clearly different patterns; i.e. the pepsin showed higher activity when the periods of larva (4∼5 ㎜TL) and juvenile II (19∼24 ㎜TL), however, the trypsin represented maximum activity at the stages of juvenile I (11∼16 ㎜TL) and young fish (27 ㎜TL), respectively.

적혈구 막에 관한 연구(I) Palmitoyl Carnitine에 의한 소 적혈구 막의 용해

이강순,장정순,조기승,이강석,Rhee, Kang-Soon,Chang, Chung-Soon,Cho, Key-Seung,Lee, Kang-Suk 생화학분자생물학회 1975 한국생화학회지 Vol.8 No.1

소의 적혈구막을 palmitoyl carnitine으로 처리하였을 때 단백질, 인지질 및 cholesterol이 동시에 완전히 용해되었고 palmitoyl carnitine에의 한 지질의 유리 및 micelle의 형성이 없는 것으로 보아 적혈구막은 lipoprotein 형태로 용해됨을 알았다. 소 적혈구막의 용해는 palmitoyl carnitine이 막의 lipoprotein과의 친수성 결합과 소수성 결합을 하므로써 용해되며 용해도는 소수성 결합 정도에의 존한다고 본다. Palmitoyl carnitine에의 한 혈구막 용해에 있어서 ATPase($Na^+$, $K^+$ dependent)와 glucose-6-phosphatase의 활성도는 저해 받지 않았으며 오히려 palmitoyl carnitine의 농도에 따라 각각 28-43%, 56-110%가 증가하였다. 용해된 막단백질에 있어 pH에 따른 용해도의 상태는 pH 5.0에서 92%의 단백질이 침전되었다. 용해된 막단백질을 Tris-glycine buffer, pH 8.3로 polyacrylamide gel electrophoresis를 행하였을 때 분자량이 450,000-70,000인 8개의 band로 분리되었다. The mechanism of solubilization of the ox-erythrocyte membrane was investigated with palmitoyl carnitine. When the membrane was treated with palmitoyl carnitine, the membrane protein, phospholipids and cholesterol were completely solubilized. It was confirmed that lipids were not liberated from membrane lipoprotein and not form micelles with palmitoyl carnitine. As a consequence, membrane was solubilized as a form of lipoprotein. In the present study, the results indicated that the erythrocyte membrane was solubilized by involvement of ionic and hydrophobic interaction of membrane lipoprotein with palmitoyl carnitine. The solubility of erythrocyte membrane depended upon the degree of the hydrophobic binding with palmitoyl carnitine. In the complete solubilized membrane, the activities of ATPase($Na^+$, $K^+$-dependent) and glucose-6-phosphatase were not affected or rather slightly increased of 28% and 56% as compared with intact membrane, respectively. The effect of pH on solubilized membrane protein showed that 92% of protein was precipitated at pH 5.0. In polyacrylamide gel electrophoresis with Tris-glycine buffer, pH 8.3, solubilized membrane protein was separated into 8 bands with molecular weight ranged from 450, 000 to 70, 000.

Swamp eel ( Monopterus albus ) 혈장 Albumin 분획의 정제 및 물리화학적 성상에 관한 연구

김상복,장정순 ( Sang Bog Kim,Chung Soon Chang ) 생화학분자생물학회 1982 BMB Reports Vol.15 No.4

Albumin fraction was purified from the swamp eel by salting out with ammonium sulfate and following by DEAE-Sephadex A50 column chromatography. The purified albumin fraction appeared to be homogeneous by Davis and NaDodSO₄ polyacrylamide gel electrophoresis after albumin fraction was treated with organic cation, rivanol (2-ethoxy-6, 9, diaminoacridine lactate). The physico-chemical properties on purified albumin fraction were characterized by means of molecular weight determination, -SH content, extinction and sedimentation coefficient, etc.