고상법에 의한 Bradykinin 유사물의 합성 (II)

최청,이재성,배만종,윤상홍,Choi, Cheong,Lee, Jae-Sung,Bae, Man-Jong,Yoon, Sang-Hong 생화학분자생물학회 1982 한국생화학회지 Vol.15 No.4

Bradykinin(BK)의 receptor $B_1$ 상에서 nonapeptide의 구조 및 활성에 대한 연구 목적으로 bradykinin유도체 ($Leu^1$, $Leu^2$)BK, ($Leu^2$, $Leu^3$)BK, ($Ala^7$, $Ala^9$)BK 및 ($Ala^8$, $Ala^9$)BK을 고상법으로 합성하였다. coupling은 N,N'-dicyclohexylcarbodiimide로 행하였으며 HBr응액으로 cleavage한 후 peptides는 Sephadex G-25와 carboxymethyl cellulose칼럼 크로마토 그라피로 정제하였다. 이들 peptides의 순도측청은 paper chromatography, thin layer chromatography, paper electrophoresis, slab gel electrophoresis. melting point 및 아미노산 분석에 따랐다. endopeptidase인 ${\alpha}$-chymotrypsin과 trypsin, exopeptidase인 carboxypeptidase A와 leucine aminopeptidase를 사용하여 이들 peptides의 분해실험을 하였다. 즉 carboxypeptidase A와 ${\alpha}$-chymotrypsin은 이들을 빠르게 분해하였고 trypsin은 느리게 분해 하였으나, ($Ala^7$, $Ala^9$) 및 ($Ala_8$, $Ala^9$)BK은 N-말단에 모두 proline의 imino와 결합되었기 때문에 leucine aminopeptidase의 작응을 받지 않았고 ($Leu^1$, $Leu^2$)BK은 1번 위치에 arginine과 같은 방향족 아미노산이 치환되지 않았으므로 trypsin의 작응을 받지 않았다. We have synthesized, by solid phase method, a series of 4 analogues of the nonapeptide, bradykinin(BK) in order to conduct a structure-activity study of these peptides on the newly discovered bradykinin $B_1$ receptor. Coupling were performed by dicyclohexylcarbodimide. After cleavage with liquid HBr, peptides were purified by gel fitration on Sephadex G-25 and ion-exchange chromatography on carboxymethyl cellulose. The purity of the nonapeptide was then checked by paper chromatography, thin layer chromatography, paper electrophresis slab gel electrophoresis, melting point and amino acid analysis. ($Leu^1$, $Leu^2$), ($Leu^2$, $Leu^3$), ($Leu^7$, $Leu^9$) and ($Leu^8$, $Leu^9$) analogues of bradykinin were incubated in vitro with endopeptidase ($\alpha$-chymotrypsin, trypsin) and exopeptidase (carboxypeptidase A, leucine aminopeptidase) in order to study the degradation patterns of peptides. ($Leu^1$, $Leu^2$), ($Leu^2$, $Leu^3$), ($Leu^7$, $Leu^9$) and ($Leu^8$, $Leu^9$) analogues of bradykinin were rapidly degraded by $\alpha$-chymotrypsin and carboxypeptidase A, while the degradation by trypsin was slow. But ($Ala^7$, $Ala^9$) BK and ($Ala^8$, $Ala^9$)BK contain imino peptide bond from proline at N-terminal and therefore they were not attacked by leucine aminopeptidase.

고상법에 의한 Bradykinin 유사물의 합성

최정,이재성,최무영,배만종 ( Cheong Choi,Jae - Sung Lee,Moo Yung Choi,Man Jong Bae ) 생화학분자생물학회 1981 BMB Reports Vol.14 No.4

Synthesis of bradykinin, (Leu¹), (Leu²) and (Ala¹, Ala^9) analogues of bradykinin by solid phase method using an improved reaction vessel was carried out. We employed gel chromatography, paper chromatography, thin layer chromatography, paper electrophoresis, disc electrophoresis and amino acid analysis for purification and for determining the purity of synthesized peptides. Bradykinin, (Leu¹), (Leu²) and (Ala¹, Ala^9) analogues of bradykinin were incubated in vitro with endopeptidases (α-chymotrypsin, trypsin) and exopeptidase (carboxypeptidase A, leucine aminopeptidase) in girder to study the degradation patterns of the peptides. Eradykinin and (Leu¹), (Leu²) and (Ala¹, Ala^9) analogues of bradykinin were rapidly degradated by α-chymotryFsin and carboxypeptidase A, while the degradation by trypsin is slow. Bradykinin, (Leu¹) and (Ala¹, Ala^9) analogues of bradykinin contain immino peptide bond from proline at N-terminal and therefore they were not attacked by leucine aminopeptidase.

고상법에 의한 Bradykinin 유사물의 합성

최청,이재성,최무영,배만종,Choi, Cheong,Lee, Jae-Sung,Choi, Moo-Yung,Bae, Man-Jong 생화학분자생물학회 1981 한국생화학회지 Vol.14 No.4

고상법으로 개량된 기구에 의하여 bradykinin, $(Leu^1)$, $(Leu^2)$ 및 ($Ala^1$, $Ala^9$) bradykinin을 합성하였으며 이들 peptide의 정제 및 순도 측정에는 gel chromatography, paper chromatography, thin layer chromatography, paper electrophoresis, disc electrophoresis 및 아미노산 분석에 의거하였다. Endopeptidase인 ${\alpha}$-chymotrypsin과 trypsin, exopeptidase인 carboxypeptidase A와 leucine aminopeptidase를 사용하여 in vitro 상에서 이들 peptide의 분해 실험을 됐다. ${\alpha}$-Chymotrypsin과 carboxypeptidase는 peptide를 빠르게 분해하였으나 trypsin의 분해 작용은 느린 것으로 나타났다. Bradykinin, $(Leu^1)$과 ($Ala^1$, $Ala^9$) bradykinin에 는 N-말단에 모두 proline의 immino기와 결합되어 있기 때문에 leucine aminopeptidase의 작용을 받지 않았다. Synthesis of bradykinin, $(Leu^1)$, $(Leu^2)$ and ($Ala^1$, $Ala^9$) analogues of bradykinin by solid phase method using an improved reaction vessel was carried out. We employed gel chromatography, paper chromatography, thin layer chromatography, paper electrophoresis, disc electrophoresis and amino acid analysis for purification and for determining the purity of synthesized peptides. Bradykinin, $(Leu^1)$, $(Leu^2)$ and ($Ala^1$, $Ala^9$) analogues of bradykinin were incubated in vitro with endopeptidases (${\alpha}$-chymotrypsin, trypsin) and exopeptidase (carboxypeptidase A, leucine aminopeptidase) in girder to study the degradation patterns of the peptides. Eradykinin and $(Leu^1)$, $(Leu^2)$ and ($Ala^1$, $Ala^9$) analogues of bradykinin were rapidly degradated by ${\alpha}$-chymotrypsin and carboxypeptidase A, while the degradation by trypsin is slow. Bradykinin, $(Leu^1)$ and ($Ala^1$, $Ala^9$) analogues of bradykinin contain immino peptide bond from proline at N-terminal and therefore they were not attacked by leucine aminopeptidase.

고상법에 의한 Bradykinin 유사물의 합성 ( 2 )

최정,이재성,배만종,윤상홍 ( Cheong Choi,Jae Sung Lee,Man Jong Bae,Sang Hong Yoon ) 생화학분자생물학회 1982 BMB Reports Vol.15 No.4

We have synthesized, by solid phase method, a series of 4 analogues of the nonapeptide, bradykinin(BK) in order to conduct a structure-activity study of these peptides on the newly discovered bradykinin Bl receptor. Coupling were performed by dicyclohexylcarbodimide. After cleavage with liquid HBr, peptides were purified by gel fitration on Sephadex G-25 and ion-exchange chromatography on carboxymethyl cellulose. The purity of the nonapeptide was then checked by paper chromatography, thin layer chromatography, paper electrophresis slab gel electrophoresis, melting point and amino acid analysis. (Leu¹, Leu²), (Leu², Leu³), (Ala^7, Ala^9) and (Ala^8, Ala^9) analogues cf bradykinin were incubated in vitro with endopeptidase (a-chymotrypsin, trypsin) and exopeptidase (carboxypeptidase A, leucine aminopeptidase) in order to study the degradation patterns of peptides. (Leu¹, Leu²), (Leu², Leu³), (Ala^7, Ala^9) and (Ala^8, Ala^9) analogues of bradykinin were rapidly degraded by a-chymotrypsin and carboxypeptidase A, while the degradation by trypsin was slow. But (Ala^7, Ala^9)BK and (Ala^8, Ala^9)BK contain imino peptide bond from proline at N-terminal and therefore they were not attacked by leucine aminopeptidase.

고등균류 균사체의 면역조절 기능성에 관한 연구

배만종,박무희,이재성 ( Man Jong Bae,Mu Hee Park,Jae Sung Lee ) 한국균학회 1996 韓國菌學會誌 Vol.24 No.2

Sarcoma-180 세포를 이용한 고등균류의 항암효과

배만종,이재성,김선희,박무희 경산대학교 생명자원개발연구소 1998 생명자원과 산업 Vol.2 No.-

상황버섯과 버들송이 균사체를 중심으로 하여 이들 균사체로부터 열수 추출된 세포내다당류와 배양액에서 회수된 다당류를 이용하여 sarcoma180에 대한 항암효과를 실험한 결과는 다음과 같다. Sarcoma-180 cell-line으로 복수임을 유발시킨 mice에 균사체와 배양액으로부터 회수한 다당체를 50㎍/㎏ 투여시의 체중증가량은 대조군에 비하여, 현저하게 체중증가량이 감소하였으며, 균사체에서 보다 배양액에서의 다당류에서 더욱 효과 있는 것으로 나타났다. 수명연장 효과를 살펴본 결과는 대조군에 비하여 상황과 버들송이 균사체의 세포외단백다당체에서 각각 47.8%, 69.6%, 32.2% 및 23.9%의 생존율을 나타내었다. In vitro에서의 S-180에 대한 증식 억제효과는 대조군에 비하여 모든 군에서 억제효과가 나타났으며, 시료의 농도에 비례하여 억제되는 것으로 나타났다. This study examine antitumor effects, the intracellular and extracellular, extracts from the liquid culture of the Phellinius igniarius and the Agrocybe cylindracea by using sarcoma180 tumor cells. The experimented was carred out by mice body weight increation, the life prolongation and tumor cell groth inhibition in vitro. The resultes obtained from the experiment as follows; Total body weight of mice, inoculated with sarcoma-180ascites tumor cells challenge, were decreased significantly in experimental groups, intracellular and extracellular polysaccharide extracts from the liquid culture of the Phellinius igniarius and the Agrocybe cylindracea than control groups. The life prolongation effects were 47.8%, 69.6%, 32.2% and 23.9%, respectively when intracellular and extracellular, extracts from the liquid culture of the Phellinius igniarius and the Agrocy cylindracea were administrated to the mice. Inhibitory efect on the growth rate of the sarcoma-180 was stronger in intracellular and extracellular, extracts from the liquid culture of the Phellinius igniarius and the Agrocybe cylindracea than control group. The growth rates of the sarcoma-180 in medium containing samples were inhibited gradually to a significant degree in proportion to the increase of the samples concentration in vitro.

고등균류의 CYTKINE 분비 및 세포증식에 미치는 영향

배만종,최청,박무희,이재성 경산대학교 생명자원개발연구소 1998 생명자원과 산업 Vol.2 No.-

담자균 유래의 신규 생리활성 물질의 탐색의 일환으로 상황버섯과 버들송이의 균사체로부터 세포내단백다당류와 세퐁외단백다당류로 분리 및 정제된 시료를 가지고 세포증식, IL-2 alc IL-4 생성의 경향에 대한 실험을 행한 결과는 다음과 같다. 1. DNA 합성과 MTT에 의한 세포증식의 결과에서는, 상황버섯과 버들송이 세포배양외액에서 추출·분리된 땅류가 균사체에서 보다 비장세포에 대한 세포활성이 높은 것으로 났다. 2. IL-2 생성 결과에서는 상황과 버들송이 균사체 및 세포외액으로부터 추출된 다당류의 농도가 10∼300㎍/mL의 범위에서 IL-2 생성이 증가되었으며, 균사체 배양외액으로부터 분리된 다당류에서 높으로 것으로 나타났다. 3. IL-4 생성의 결과는 상황버섯과 버들송이로부터 추출된 세포내 및 세포외 단백다당류가 대조군에 비행서 높은 것으로 나타났다. The polysaccharide, intracellurar and extracellular, extracts from the liquid culture of the Phellinius igniarius and the Agrocybe cylindracea were purifide. This study was conducted to investigated which were measured the proliferative response to mitogens of splenocyte and production of IL-2 and IL-4. The results obtained from the experiment as follows : The proliferation of splenocyte was increased mosteffectively when the extracellua polysaccharide extracted from the Phellinius igniarius and the Agrocybe cylindracea were administrated to the mice. And the extractcellular polysaccharide extract from the Phellinius igniarius was proliferation activity increased according to culture time. The IL-2 secreased, a marker increased when 48hrs. culture with the extracellula polysaccharide extracted from the Phellinius igniarius and the Agrocybe culindracea(10∼100㎍/mL). And there was significantly the extracellula polysacchride extracted from the Phellinius then the extracellula polysaccharide extracted from the Agrocybe cylindracea. IL-4 production was significantly increased compared with the control group.