Study on the Distribution and Synthesis of Apolipoprotein B Subspecies in Rat Lymph Chylomicrons and Plasma Lipoproteins

함경수,이영식,이남진,Hahm, Kyung-Soo,Lee, Young-Sick,Lee, Nam-Jeen 생화학분자생물학회 1985 한국생화학회지 Vol.18 No.1

백서의 intestinal lymph에서 lymph chylomicrons를 분리하였으며, 백서의 post-heparin plasma와 functionally hepatectomize시키고 eviscerate시킨 백서로 부터 각각 chylomicron remnants를 형성시켜 분리하였다. 이 chylomicrons와 plasma liporpoteins를 4% SDS-PAGE로 분석하여, apo B subspecies의 분표를 확인하고 human lipoproteins와 비교하였으며 그 결과에 의하면 백서의 plasma lipoproteins(VLDL, IDL, LDL)는 human과 달리 apo B-100와 B-48만 함유하고 있으며, 이 중 apo B-48은 백서의 intestine에서 합성되어 chylomicrons로 분비되는 유일한 apo B subspecies임을 밝혔다. Rat lymph chylomicrons were prepared by using post-heparin plasma and from the functionally hepatectomized and eviscerated rats. Rat chylomicrons and plasma lipoproteins were analyzed by 4% SDS-PAGE for the distribution of apo B subspecies and compared with that of human. The result showed that rat plasma lipoproteins (VLDL, IDL, and LDL) all contain both apo B-48 which differed from human lipoproteins. The result also provided an evidence that rat apo B-48 is the only apo B subspecies synthesized in the intestine and secreted in the chylomicrons.

혈장 저밀도 지단백질 : 저밀도 지단백질에 대한 모노크로날 항체를 이용한 아포지단백질 B 의 구조

함경수 ( Kyung Soo Hahm ) 한국생화학분자생물학회 (구 한국생화학회) 1986 생화학총설집 Vol.1 No.-

담석에서 추출한 단백질의 분석

함준수(Joon Soo Hahm),박경근(Kyung Geun Park),박준용(Joon Yong Park),한동수(Dong Soo Han),이민호(Min Ho Lee),기춘석(Choon Suhk Kee),박경남(Kyung Nam Park),이광수(Kwang Soo Lee),최은아(Eun A Choi),이명규(Myung Kyo Lee),함경수(Kyung So 대한소화기학회 1996 대한소화기학회지 Vol.28 No.1

N/A Background/Aims: Biliary proteins have been suggested to play an important role in nucleation and gallstone formation. However, the exact roles and characteristics have not been completely documented. The aim of the present study is to isolate and characterize the nucleating protein extracted from gallstones. Methods: We tried to extract, isolate and characterize proteins in patients with gallstones. Twenty-two gallstones were obtained(12 cholesterol, 10 pigment) at cholecystec- tomy and extracted with ethanol/ether mixture. Then, isoelectric focusing was performed and gallstone proteins were analysed by SDS-polyacrylamide gel electrophoresis. The amino acids were also analysis by the autoamimo acid analyzer system. Results: The mean amount of gallstone protein was 4.15 mg/(g stone) in cholesterol and 16.15 mg/(g stone) in pigment stone. The proteins from both cholesterol and pigment stones showed major bands at low pH on isoelectric focusing. On SDS-PAGE, low molecular protein bands were noted, mainly below 45KD in both cholesterol and pigment stones. The composition of aspartate and glutamate was 21.6% in cholesterol stones and 22.7% in pigment stones. Conclusions: The proteins in cholesterol and pigment gallstones are low molecular weight acidic proteins, and these acidic proteins seem to play an important role in the pathogenesis of gall stones. However, it remains to be determined whether these proteins differ in functional roles from different gallstones. (Korean J Gastroenterol 1996;28: 92 - 100)

RID에 의한 혈장 Apolipoprotein

이명규,함경수 ( Myeong Kyu Lee,Kyung Soo Hahm ) 생화학분자생물학회 1986 BMB Reports Vol.19 No.4

Radial immunodiffusion (RID) was used for determination of apolipoprotein B in LDL, VLDL, and plasma. The method closely approximated the values of LDL apo B obtained by the standard Lowry procedure (r=0.968). Using 1% agarose in the method made it feasible to estimate plasma and VLDL apo B levels. The present study also shows that plasma total cholesterol level correlates well (r=0.981) with the plasma apo B level measured by the RID method.

Quantitation of Human Plasma Apoprotein B by Radial Immunodiffusion

이명규,함경수,Lee, Myeong-Kyu,Hahm, Kyung-Soo 생화학분자생물학회 1986 한국생화학회지 Vol.19 No.4

Radial immunodiffusion (RID)방법을 이용하여 LDL, VLDL 및 혈장에 존재하는 apoliporprotein B의 농도를 측정하였다. LDL apo B 의 농도를 본 방법과 단백질 정량 방법으로 흔히 사용되는 Lowry의 방법으로 측정, 비교하여 보았을 때 차이가 없었다 (r=0.968). Agarose 의 농도를 1%로 함으로써 VLDL 과 혈장의 apo B의 농도 또한 측정이 가능하였으며, 본 방법으로 측정한 혈장의 apo B 농도와 혈장 콜레스테롤 농도를 비교해 보았을 때 의의있는 상관관계(r=0.981)가 있었다. Radial immunodiffusion (RID) was used for determination of apolipoprotein B in LDL, VLDL, and plasma. The method closely approximated the values of LDL apo B obtained by the standard Lowry procedure (r=0.968). Using 1% agarose in the method made it feasible to estimate plasma and VLDL apo B levels. The present study also shows that plasma total cholesterol level correlates well (r=0.981) with the plasma apo B level measured by the RID method.

인체 간세포 표면에 존재하는 쥐의 간암의 특이 항원에 대한 면역학적 규명

이명규,한문희,함경수 ( Myeong Kyu Lee,Moon H . Han,Kyung Soo Hahm ) 생화학분자생물학회 1988 BMB Reports Vol.21 No.4

Hepatocellular carcinoma was induced in Sprague-Dawley rats using 2-MeDAB and DENA, and hepatoma associated antigens were identified and isolated from the plasma membrane surface of rat liver. Rabbit antisera against hepatoma associated antigens were obtained and reactivity of the antibody with proteins extracted from human livers (normal and hepatoma) was analyzed by double immunodiffusion, immunoelectrophoresis and crossed-immunoelectrophoresis. The results showed a strong evidence for the presence of proteins (MW 65,000 & 55,000) on the human liver cell cross-reacting with the antibody against rat hepatoma associated antigens. These human proteins were found to be unreactive with the antiserum against plasma membrane surface proteins of normal rar liver.

Immunoaffinity Purification and Receptor Assay for Polymerized Human Serum Albumin of Pre-S2 Peptide on Hepatitis B Virus

윤혜영,한문희,함경수,Yun, Hye-Young,Han, Moon-H.,Hahm, Kyung-Soo 생화학분자생물학회 1988 한국생화학회지 Vol.21 No.4

대장균내에서 과량 발현된 재조합 프리-S2 펩티드를 면역친화력 크로마토그래피에 의해 분리 정제하였으며, 이 펩티드에 대한 중합 인혈청 알부민(pHSA)의 결합을 연구하였다. 프리-S2 부위의 pHSA수용체 활성을 검출하는 방법을 프리-S2-$\beta$ 갈락토시다제 융합단백질을 이용하여 개발하였으며, 민감도는 10-100 p mol의 프리-S2에 해당되었다. 또한 이 방법은 프리-S2 농도 결정 뿐 아니라 pHSA수용체 활성을 중화시킬 수 있는 인자들을 측정하는데 이용될 수 있다. Recombinant pre-S2 peptide was purified to apparent homogeneity from E. coli by immunoaffinity chromatography using monoclonal antibody and studied for its binding characteristics with monoclonal antibody and pHSA (polymerized human serum albumin). A simple and sensitive assay method for detecting the activity of pHSA receptor on pre-S2 peptide of hepatitis B virus surface antigen (HBsAg) has been developed using pre-S2-$\beta$-galactosidase fusion protein. The assay can be used for determining pre-S2 or factors neutralizing pHSA receptor activity. The sensitivity of the assay is about 10 to 100 pmol of pre-S2.

B형 간염 바이러스 프리 - S2 펩티드의 분리 정제 및 중합 인혈청 알부민 수용체 검출법

윤혜영,한문희,함경수 ( Hye Young Yun,Moon H . Han,Kyung Soo Hahm ) 생화학분자생물학회 1988 BMB Reports Vol.21 No.4

Recombinant pre-S2 peptide was purified to apparent homogeneity from E. coli by immunoaffinity chromatography using monoclonal antibody and studied for its binding characteristics with monoclonal antibody and pHSA (polymerized human serum albumin). A simple and sensitive assay method for detecting the activity of pHSA receptor on pre-S2 peptide of hepatitis B virus surface antigen (HBsAg) has been developed using pre-S2-β-galactosidase fusion protein. The assay can be used for determining pre-S2 or factors neutralizing pHSA receptor activity. The sensitivity of the assay is about 10 to 100 pmol of pre-S2.

Immunological Identification of Rat Hepatoma Associated Antigens on Human Liver

이명규,한문희,함경수,Lee, Myeong-Kyu,Han, Moon-H.,Hahm, Kyung-Soo 생화학분자생물학회 1988 한국생화학회지 Vol.21 No.4

화학 발암제인 2-MeDAB 및 DENA를 투여함으로서 Sprague-Dawley 쥐에 간암을 유발시켰으며, 간암세포 원형질막 표면으로부터 간암 특이 항원을 분리하였다. 쥐의 간암 특이 항원에 대한 항체를 토끼로부터 분리하였으며, 이 항체와 사람의 정상간 및 간암세포 표면에서 분리한 단백질과의 반응을 double immunodiffusion, 면역 전기영동법 및 교차변역 전기영동법 등을 이용하여 분석하였다. 그 결과 사람의 간 세포 원형질막 표면에 쥐의 간암 특이 항원에 특이한 항체와 반응하는 단백질(분자량 65,000 및 55,000)이 존재함을 확인하였다. 한편 이 단백질은 쥐의 정상 간세포 표면 단백질에 대한 항체와는 반응하지 않았다. Hepatocellular carcinoma was induced in Sprague-Dawley rats using 2-MeDAB and DENA, and hepatoma associated antigens were identified and isolated from the plasma membrane surface of rat liver. Rabbit antisera against hepatoma associated antigens were obtained and reactivity of the antibody with proteins extracted from human livers (normal and hepatoma) was analyzed by double immunodiffusion, immunoelectrophoresis and crossed-immunoelectrophoresis. The results showed a strong evidence for the presence of proteins (MW 65,000 & 55,000) on the human liver cell cross-reacting with the antibody against rat hepatoma associated antigens. These human proteins were found to be unreactive with the antiserum against plasma membrane surface proteins of normal rar liver.

Recombinant Human Interleukin-2: II. Biological and Therapeutic Activities

최혜림,윤혜영,함경수,Choi, Hye-Lim,Yun, Hye-Young,Talmadge, James E.,Hahm, Kyung-Soo 생화학분자생물학회 1988 한국생화학회지 Vol.21 No.2

Recombinant human interleukin-2 (rH IL-2, $Ser^{125}$-rH IL-2) purified from E. coli was characterized its biological and therapeutic activities for the establishment of preclinical screening system and therapeutic applications. In vitro augmentation of natural killer (NK) cell activity and mixed lymphocyte reaction (MLR) assay were shown to be comparable with other rH IL-2 from different manufacturers and laboratories. An antiserum against rH IL-2 was obtained from chick, and characterized the reacticvity with rH IL-2 reconstituted from lyophilized state in relation to the stability and conformational changes of the recombinant protein. In addition, long term storage of rH IL-2 in lyophilized or solution was not shown to incur remarkable loss of MLR activities, and any appearance of either degradation or aggregation products when analyzed by SDS-PAGE, indicating the stability and suitability for therapeutic applications of rH IL-2. 대장균으로 부터 분리정제된 재조합 인체 인터루킨-2 (KAIST rH IL-2, $Ser^{125}$-rH IL-2)를 전임상시험 및 암치료에 응용하기 위한 목적으로 생물학적 및 면역요법적 활성도를 측정하였다. In vitro에서 자연살해세포 활성의 증진능력 및 혼합 임파구반응 실험에서 활성을 나타냈으며 다른 회사의 재조합 인터루킨-2 제품들과 유사한 결과를 얻었다. 재조합 인체 인터루킨-2의 입체구조적 및 생물학적 활성도의 안정성을 조사하기 위하여 정제된 재조합 인체 인터루킨-2에 대한 항혈청을 닭에서 얻었으며 동결건조 저장하였던 재조합 인체 인터루킨-2와 이 항체와의 반응을 조사하였으며, 또한 6개월간 저장하였던 인터루킨-2의 안정성을 혼합 임파구반응 및 전기영동 방법으로 그 활성 및 단백질 성분을 조사한바 혼합임파구 반응에서 여전히 활성을 보여주였으며, 전기영동상 분해되었거나 응집된 산물이 생기지 않았음을 확인하였다. 이상의 결과는 KAIST 재조합 인체 인터루킨-2가 전임상시험 및 임상시험 목적에 사용하기에 적합함을 시사한다.